Objective To investigate the prevalence of mutation in the locus 306 of embB gene in multi-drug resistant (MDR) Mycobacterium tuberculosis (TB) and evaluate the prospects for using it as a molecular marker to detect MDR-TB.Methods The 291 strains enrolled in this study were from the reference laboratory of Shanghai municipal centers for disease control and prevention, all of which had been tested for drug susceptibility.Mutation in embB 306 was screened both by amplification refractory mutation system (ARMS) and DNA sequencing.The mutation frequencies of embB 306 in the sample groups varied in drug resistance were statistically analyzed.Results 38(51.4% ) of the 74 MDR-TB were embB 306-mutant (X2 =93.8,P＜0.01).Of the 24 TB resistant to at least two drugs but not MDR, 9(37.5% ) were embB 306 mutant (X2 =60.1 ,P＜0.01 ).But only two(4.9% ) embB 306-mutant strains were found in 41 strains resistant to only one drug (X2 =6.8,P=0.0093).None embB 306-mutant strains were found in 152 pansensitive strains.The specificity of using embB 306 as a molecular marker for detecting multi-drug resistant TB was 94.9% (206/217).Conclusions As a molecular marker for screening drug resistant TB,especially MDR-TB, the gene locus embB 306 shows a relatively high sensitivity and specificity, promising a sound future for its application in clinics to realize fast screening of patients infected with MDR-TB and to provide evidence for appropriate medication.

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.

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.