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Objective@#To investigate expressions of Toll-like receptor(TLR)-2, TLR-4 and TLR-6 in peripheral blood mononuclear cells (PBMC) and serum immunoglobulin G (IgG) and IgM levels in pediatric idiopathic nephrotic syndrome (INS). The correlation between Toll-like receptors (TLR-2, TLR-4 and TLR-6) and serum immunoglobulin levels (IgG and IgM) will be proven in the pathogenesis of childhood INS in active stage (AS) and remission stage (RS).@*Methods@#Forty-two INS patients (experimental group, 32 boys, 10 girls) and 29 healthy individuals (healthy control group, 19 boys, 10 girls) were enrolled in the present study from June, 2017 to October, 2018 in the First Affiliated Hospital of Xinxiang Medical University. There were no significant differences in gender (χ2=0.966, P=0.326) and age (t=-0.139, P=0.89) between 2 groups.TLR-2 mRNA, TLR-4 mRNA and TLR-6 mRNA expressions of PBMC were evaluated by using real-time PCR in both INS children with AS and RS and healthy children.The blood samples were drawn to detect by way of immunoturbidimetry serum immunoglobulin (IgG and IgM) levels in both INS children with AS and healthy children.The correlation between Toll-like receptors (TLR-2 mRNA, TLR-4 mRNA, TLR-6 mRNA) and serum immunoglobulin (IgG, IgM) levels were analyzed by using spearman correlation analysis.@*Results@#The expression levels of TLR-2 mRNA in INS children with AS, RS and healthy children were 1.85±0.30, 1.00±0.23 and 0.85±0.12, respectively, and the expression levels of TLR-4 mRNA were 1.24±0.27, 0.80±0.23 and 0.68±0.09, respectively, while the expression levels of TLR-6 mRNA were 1.35±0.23, 0.92±0.19 and 0.79±0.15, respectively, so the differences were statistically significant(F=198.453, 67.013, 82.405, all P<0.01). The serum IgG level(2.90±0.89) g/L was lower in INS children with AS compared with the controls(9.21±1.88) g/L.However, the serum IgM level (2.87±0.96) g/L was higher in INS children with AS when compared with the controls(1.48±0.30), and the differences were statistically significant(t=-16.810, 8.701, all P<0.05). No significant correlation was noted between Toll-like receptors(TLR-2 mRNA, TLR-4 mRNA, TLR-6 mRNA) and serum immunoglobulin levels (IgG, IgM) (all P>0.05).@*Conclusions@#The results indicate that Toll-like receptors (TLR-2, TLR-4 and TLR-6) and immunoglobulin (IgG, IgM) might play a role in the pathogenesis of pediatric INS via distinct pathway.
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Objective To investigate expressions of Toll-like receptor (TLR)-2,TLR-4 and TLR-6 in peripheral blood mononuclear cells (PBMC) and serum immunoglobulin G (IgG) and IgM levels in pediatric idiopathic nephrotic syndrome (INS).The correlation between Toll-like receptors (TLR-2,TLR-4 and TLR-6) and serum immunoglobulin levels (IgG and IgM) will be proven in the pathogenesis of childhood INS in active stage (AS) and remission stage (RS).Methods Forty-two INS patients (experimental group,32 boys,10 girls) and 29 healthy individuals (healthy control group,19 boys,10 girls) were enrolled in the present study from June,2017 to October,2018 in the First Affiliated Hospital of Xinxiang Medical University.There were no significant differences in gender (x2 =0.966,P =0.326) and age (t =-0.139,P =0.89) between 2 groups.TLR-2 mRNA,TLR-4 mRNA and TLR-6 mRNA expressions of PBMC were evaluated by using real-time PCR in both INS children with AS and RS and healthy children.The blood samples were drawn to detect by way of immunoturbidimetry serum immunoglobulin (IgG and IgM) levels in both INS children with AS and healthy children.The correlation between Toll-like receptors (TLR-2 mRNA,TLR-4 mRNA,TLR-6 mRNA) and serum immunoglobulin (IgG,IgM) levels were analyzed by using spearman correlation analysis.Results The expression levels of TLR-2 mRNA in INS children with AS,RS and healthy children were 1.85 ± 0.30,1.00 ± 0.23 and 0.85 ± 0.12,respectively,and the expression levels of TLR-4 mRNA were 1.24 ± 0.27,0.80 ± 0.23 and 0.68 ± 0.09,respectively,while the expression levels of TLR-6 mRNA were 1.35 ± 0.23,0.92 ± 0.19 and 0.79 ± 0.15,respectively,so the differences were statistically significant (F =198.453,67.013,82.405,all P ＜ 0.01).The serum IgG level(2.90 ± 0.89) g/L was lower in INS children with AS compared with the controls (9.21 ± 1.88) g/L.However,the serum IgM level (2.87 ± 0.96) g/L was higher in INS children with AS when compared with the controls(1.48 ± 0.30),and the differences were statistically significant(t =-16.810,8.701,all P ＜0.05).No significant correlation was noted between Toll-like receptors (TLR-2 mRNA,TLR-4 mRNA,TLR-6 mRNA) and serum immunoglobulin levels (IgG,IgM) (all P ＞ 0.05).Conclusions The results indicate that Toll-like receptors (TLR-2,TLR-4 and TLR-6) and immunoglobulin (IgG,IgM) might play a role in the pathogenesis of pediatric INS via distinct pathway.
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Objective To investigate the expressions of Toll-like receptor (TLR)-1,TLR-3,TLR-9 in peripheral blood mononuclear cells (PBMC) of children with idiopathic nephrotic syndrome (INS).Methods The expressions of TLR-1,TLR-3 and TLR-9 mRNA were measured by real time-polymerase chain reaction (RT-PCR) in PBMC of 45 children with INS came from the First Affiliated Hospital of Xinxiang Medical University before and after treatment (active group and remission group,respectively) and 30 healthy children from medical examination center as healthy control group,so as to explore the relativity between TLR-1,TLR-3 and TLR-9 mRNA and serum levels of albumin and cholesterol.Results The expression of TLR-1 mRNA/β-actin in PBMC of children with INS before treatment (the active group) was significantly higher than that in the remission group (2.432 ± 0.231 vs.1.675 ± 0.627,t =7.599,P ＜ 0.05) and the healthy control group (2.432 ± 0.231 vs.0.512 ± 0.228,t =35.446,P ＜0.05),and the differences were statistically significant in the expression of TLR-1 mRNA/β-actin between remission group and healthy control group (1.675 ± 0.627 vs.0.512 ± 0.228,t =9.722,P ＜ 0.05).The expression of TLR-3 mRNA/β-actin in PBMC of children with INS before treatment (the active group) (0.987 ± 0.124) was significantly higher than that in the remission group (0.501 ± 0.016) and the healthy control group (0.021 ± 0.001),the differences were statistically significant (t =26.076,42.571,all P ＜ 0.05),and the remission group was significantly higher than that in healthy control group (t =163.732,P ＜ 0.05).The expression of TLR-9 mRNA/β-actin in PBMC of children with INS before treatment (activity group) (1.965 ±0.952) was significantly higher than that in the remission group (1.336 ±0.282) and the healthy control group (0.790 ±0.731),the differences were statistically significant (t =4.249,5.734,all P ＜ 0.05),and the remission group was significantly higher than that in healthy control group (t =4.541,P ＜ 0.05).The serum levels of albumin in children with INS before treatment (activity group) (23.62 ± 11.67) g/L was significantly lower than that in remission group (42.19 ± 16.33) g/L and healthy control group (46.88 ± 14.80) g/L,and the differences were statistically significant (t =-6.20,-7.58,all P ＜0.01).The serum levels of cholesterol in children with INS before treatment (the active group) (9.54 ±2.53) mmol/L was significantly higher than that in the remission group (3.01 ± 1.72) mmoL/L and the healthy control group (2.89 ± 1.66) mmol/L,and the differences were statistically significant (t =14.32,12.677,all P ＜ 0.01).The serum levels of albumin and cholesterol in children with INS after treatment were not statistically significant difference compared to the healthy control group (t =-1.264,0.30,all P ＞ 0.05).The correlation analysis showed that there was a negative correlation between the expressions of TLR-1,TLR-3 and TLR-9 mRNA and the serum level of albumin in PBMC of children with INS before treatment (r =-0.457,-0.891,-0.125,respectively,all P ＜ 0.05).However,there was a positive correlation between the expressions of TLR-1,TLR-3 and TLR-9 mRNA and the serum level of cholesterol (r =0.445,0.911,0.872,all P ＜ 0.05).Conclusions The expressions of TLR-1,TLR-3 and TLR-9 mRNA in PBMC of children with INS were significantly higher and correlated with activity of disease.
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OBJECTIVE: To study the variabilities of the effects of cold pathogen and cold-dampness pathogen on the fractalkine (FKN) mRNA expression in lung tissues of rats. METHODS: Twenty-four Wistar rats of SPF grade were randomly divided into 3 groups: normal temperature group, cold pathogen group and cold-dampness pathogen group. There were 8 rats in each group. Rats in normal temperature group were bred at (20+/-2)degrees centigrade and under normal humidity (50%-55%) for 2 h. Rats in cold pathogen group were bred at -10 degrees centigrade and under normal humidity (50%-55%) for 2 h, and the rats in cold-dampness pathogen group were bred at -10 degrees centigrade and under high humidity (90%-100%) for 2 h. Rats in the three groups were bred in thermostats under the corresponding conditions on the first day of experiment, and then the rats in different groups were all bred at normal temperature. Lung specimens in 3 groups were gathered four days later. The behavior and the pathological changes in the lung tissues of rats in different groups were observed. The content of interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-alpha) in lung homogenate was detected by radioimmunoassay (RIA) method. Expression of FKN mRNA in lung homogenate was detected by reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: The lung tissues of rats in both cold pathogen group and cold-dampness pathogen group had various degrees of pathological changes. Compared with normal temperature group, the content of IL-6 and TNF-alpha was increased obviously in lung homogenate of rats in both cold pathogen group and cold-dampness pathogen group (P<0.01). The content of IL-6 and TNF-alpha in lung homogenate of rats in cold-dampness pathogen group was obviously higher than that in cold pathogen group (P<0.01). The RT-PCR results showed a low expression of FKN mRNA in lung tissues of rats in normal temperature group. If the injured lung tissues were aggravated, the expression of FKN mRNA in the lung tissues was elevated. Compared with normal temperature group, FKN mRNA expressions in both cold pathogen group and cold-dampness pathogen group were increased obviously (P<0.01). FKN mRNA expression in lung homogenate of rats in cold-dampness pathogen group was also obviously higher than that in cold pathogen group (P<0.01). CONCLUSION: Cold pathogen can induce lung injury and up-regulate the FKN mRNA expression in lung tissue. Dampness pathogen can up-regulate the FKN mRNA expression through aggravating the injury of lung tissues caused by cold pathogen. FKN has a close relationship with the lung injury.
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OBJECTIVE: To explore the effects of wind and dampness pathogens on cytokines in the lung tissue of rats with cold syndrome due to different gradient cold pathogens. METHODS: One hundred and four Wistar rats of SPF grade were randomly divided into 13 groups: normal temperature group, six cold pathogen groups and six cold plus wind-dampness pathogen (wind of grade 5 and 90%-100% relative humidity) groups. The cold pathogens were constant low temperature (including 10 degrees C, 0 degree C, -10 degrees C) and temperature change (including 20 to 10 degrees C, 20 to 0 degrees C, and 20 to -10 degrees C). The rats in different groups were kept in a temperature-controlled box under the corresponding condition for 2 hours on the first day of experiment. Then, the rats were all raised in normal temperature for 4 days and the rats' behaviors were observed. The contents of tumor necrosis factor alpha (TNF-alpha), interleukin-6(IL-6) and interleukin-4 (IL-4) in lung homogenate were measured by radioimmunoassay and the content of interferon-gamma (IFN-gamma) was detected by enzyme-linked immunosorbent assay. RESULTS: In comparison with cold pathogen groups, contents of TNF-alpha, IL-6 and IL-4 were obviously increased in lung homogenate of rats in cold plus wind-dampness pathogen groups (P<0.01), and the content of IFN-gamma and IFN-gamma/IL-4 ratio were obviously decreased (P<0.01). CONCLUSION: Wind-dampness pathogen can seriously aggravate the injury to lung tissue caused by cold pathogen, and the unbalance of Th(1)/Th(2) in lung homogenate of rats.