As an important protein molecule for innate immunity,Toll-like receptor 7 (TLR7) is also a bridge between innate immunity and adaptive immunity.When TLR7 is activated,it can exert its effect on liver diseases through different signaling pathways.Studies on the role and application of TLR7 agonist in liver diseases are increasing in recent years.This article reviewed the research on TLR7 agonist in liver diseases.

Objective:To observe the protein and mRNA expressions of microtubule-associated protein 1 light chain 3 (LC3)B, P62 and Beclin1 in the liver of rats with chronic fluorosis, and to explore the role of autophagy in pathogenesis of liver injury induced by fluorosis.Methods:Using a group design, 54 SD rats were divided into 9 groups according to their weight (100 - 120 g) using a random number table method, each group with 6 rats, half male and half female. They were control group (NC group), low fluoride group (LF group), high fluoride group (HF group), NC + rapamycin (RAP) group, LF + RAP group, HF + RAP group, NC + chloroquine (CQ) group, LF + CQ group, and HF + CQ group. The NC group drank tap water (fluoride concentration was 0.5 mg/L), LF group drank fluoride water (fluoride concentration was 5.0 mg/L), HF group drank fluoride water (fluoride concentration was 50.0 mg/L); NC + RAP group, LF + RAP group and HF + RAP group were fed with corresponding drinking water, respectively, for 3 months, and then RAP (1.5 mg/kg) was intraperitoneally administered for 10 d; NC + CQ group, LF + CQ group and HF + CQ group were fed with corresponding drinking water, respectively, for 3 months, and then CQ (60 mg/kg) was intraperitoneally administered for 10 d. Bone and 24-hour urine samples of rats in each group were collected to detect the contents of bone fluoride and urine fluoride; liver histomorphological changes were observed through hematoxylineosin staining; protein and mRNA expressions of LC3B, P62 and Beclin1 in liver were detected by immunohistochemistry and real-time fluorescence quantitative PCR, respectively.Results:Compared with the NC group [(0.03 ± 0.00) mg/kg, (0.34 ± 0.08) mg/L], the contents of bone fluoride [(3.86 ± 0.08) mg/kg] and urine fluoride [(1.11 ± 0.16) mg/L] in HF group were higher ( P < 0.05). In the NC group, the lobule structure of liver tissue was clear, the hepatic cords were arranged in order, and the cell structure was normal. There were different degrees of hepatocyte edema in LF and HF groups. After intraperitoneal injection of RAP, compared with the corresponding fluoride group, the morphology of hepatocytes did not change significantly. After intraperitoneal injection of CQ, compared with the corresponding fluoride group, the liver cells showed obvious edema, and the degree of edema aggravated with the increase of fluoride concentration. Compared with the NC group, the protein expressions of LC3B and Beclin1 in HF group were higher ( P < 0.05), and the protein expression of P62 was lower ( P < 0.05). After intraperitoneal injection of RAP, the protein expressions of LC3B and P62 in LF + RAP group was lower than that in LF group ( P < 0.05); Compared with HF group, the protein expressions of LC3B and Beclin1 in HF + RAP group were lower ( P < 0.05). After intraperitoneal injection of CQ, protein expression of P62 in LF + CQ group was higher than that in LF group ( P < 0.05); Compared with HF group, protein expression of P62 in HF + CQ group was higher ( P < 0.05). Conclusions:Early (3 month) fluoride intake could promote autophagy and induce edema of hepatocytes in rats, and RAP had similar effects. CQ may induce liver injury by inhibiting autophagy of hepatocytes.

The aim of this study is to establish an HPLC method for simultaneous determinations of mifepristone and its metabolites, mono-demethylated mifepristone, di-demethylated mifepristone and C-hydroxylated mifepristone in plasma and to evaluate the pharmacokinetic characteristics of mifepristone tablet. Twenty healthy female Chinese subjects were recruited and a series of blood samples were collected before and after 0.25, 0.5, 1.0, 1.5, 2.0, 4.0, 8.0, 12.0, 24.0, 48.0, 72.0 and 96.0 hours administration by a single oral dose of 75 mg mifepristone tablet. Mifepristone and its three metabolites were extracted from plasma using ethyl acetate and determined by high performance liquid chromatography. The main pharmacokinetic parameters of mifepristone and its metabolites, including Cmax, tmax, MRT, t(1/2), V, CL, AUC(0-96 h) and AUC(0-infinity), were calculated by Drug and Statistical Software Version 2.0. The simple, accurate and stable method allows the sensitive determinations ofmifepristone and its metabolites in human plasma up to 4 days after oral administration of 75 mg mifepristone tablet and the clinical applications of their pharmacokinetic studies.

Objective To evaluate the clinical therapeutic efficacy and adverse reactions of Coix Seed Extract combined with transcatheter arterial chemoembolization(TACE) in the treatment for hepatic metastases and to compare the levels of serum VEGF of the patients before and after the treatment.Methods FromJuly 2009 to July 2011, 62 patients with hepatomatic metastases were enrolled in this study and were randomized into combining group and control group.The combining group were treated by TACE combined with Coix Seed Extract, and the control group were treated with TACE only. Both groups underwent TACE at least twice.Before and after the treatment,the enzyme linked immunosorbentassay(ELISA)was used to examine the change of VEGF. Results Among 62 assessable patients, overall response rate was 61.3 ％(19/31) in combining group and 45.2 ％ (14/31)in control group respectively (P＞ 0.05).The disease control rate was 80.6 ％(25/31) in combining group and 54.8 ％ (17/31) in control group(P＜0.05).The 1-year survival rates were 67.7 ％(21/31) in combining group and 38.7 ％(12/31) in control group(P ＜ 0.05).The median PFS(progression free survival time) was 7.0 months in combining group and 5.1 months in control group (P＜ 0.05).The levels of serum VEGF in combining group were significantly decreased after treatment (P ＜ 0.05). There was no significant difference between the levels of serum VEGF before and after treatment in control group(P＞ 0.05).The levels of serum VEGF in combining group after treatment were lower than that in control group after treatment (P ＜ 0.05). Adverse reactions in both groups were grade Ⅰ to Ⅲ.The fatigue,anorexia and the right upper quadrant pain was significantly superior in combining group. Conclusion Coix Seed Extract combined with transcatheter arterial chemoembolization in the treatment of hepatomatic metastases can improve disease control rate,decrease the levels of serum VEGF,improve clinical symptoms,prolong the PFS and 1-year survival rate.

Objective:To investigate the prevalence of autism spectrum disorders (ASD)among children in kindergartens in Xi′an urban districts.Methods:A stratified cluster sampling method was adopted, and selected all children from 12 kindergartens in 6 urban districts of Xi′an.Primary screening positive children with ASD were identified by filling out the Autism Behavior Checklist (ABC) by their parents and the guardians reports, and then the beha-vioral observations were made to identify suspicious ASD children.Finally these children were diagnosed in the hospital through the autism diagnostic observation schedule, 2 nd edition (ADOS-2). The data were calculated with SPSS 18.0. Results:Totally, 38 cases with ASD were diagnosed among 5 178 children, the prevalence of children ASD in kindergartens in Xi′an urban districts was 7.3‰, and the 95% confidence interval was 4.98‰-9.62‰.The prevalence of ASD in children was statistically significant in different age groups ( χ2=9.914, P<0.05) and gender groups ( χ2=18.812, P<0.05). The accuracy of ASD screening by guardians reports is better than that by ABC. Conclusions:(1)The prevalence of ASD children in kindergartens in Xi′an urban districts is at high level in similar reports in China.If ASD children at the special education institutions and home are considered, the overall prevalence rate is higher, indicating that the prevalence of ASD children in China may be underestimated.(2)With the increase of age, the attendance rate of ASD children in kindergartens has a decreasing trend.

Wheat grain peroxidase 1 (WP1) belonged to class III plant peroxidase with cofactor heme, which not only has antifungal activity, but also influences the processing quality of flour. In order to enhance functional expression of WP1 in prokaryotic system by increasing endogenous heme synthesis, we constructed a recombinant plasmid pACYC-A-L containing hemA and hemL of Esherichia coli. Then, we co-transformed it into host strain T7 Express with secretive expression vector (pMAL-p4x-WP1) or non-secretive expression vector (pET21a-MBP-WP1), respectively. The MBP-WP1 fusion protein was further purified by amylose affinity chromatography and its peroxidase activity was assayed using 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonate) (ABTS) as substrate. At 12 h after induction at 28 degree, the extracellular 5-aminolevulinic acid (5-ALA) production of T7 Express/pACYC-A-L was up to 146.73 mg/L, simultaneously the extracellular porphrins also increased dramatically. The peroxidase activity of functional MBP-WP1 obtained from T7 Express/ (pACYC-A-L + pMAL-p4x-WP1) was 14.6-folds of that purified from T7 Express/ pET21a-MBP-WP1. This study not only successfully enhanced functional expression of wheat peroxidase 1 in Esherichia coli, but also provided beneficial references for other important proteins with cofactor heme.
Escherichia coli ; genetics ; metabolism ; Genetic Vectors ; genetics ; Heme ; biosynthesis ; genetics ; Peroxidases ; biosynthesis ; genetics ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; Transformation, Genetic

Objective: To explore the relationship between non-suicidal self-injury and Internet addictive behavior in adolescents. Methods: A cross-sectional survey of 15 538 middle school students across 5 provinces in China. Participants reported self- injury behavior during the past year and the Internet addition behaviors were assessed through Young’s Internet Addiction Test(IAT). Results: The prevalence of self-reported 1-year NSSI, Internet addictive behavior and pre-addictive behaviors was 28.7%, 3.3% and 29.9%, respectively. After controlling for potential confounders, the Internet addition was positively correlated with NSSI, and the risk of all types of NSSI was higher in adolescent with Internet addition than those with mild suspected Internet addictive behavior(OR=1.3-2.8). Conclusion These findings indicate that Internet addition is associated with the incidence of NSSI in adolescents.

Objective: To investigate the relationship between neglect , physical abuse and non-suicidal self-injury (NSSI) behaviors among middle school students in rural China. Methods: A total of 14 162 adolescents in Anhui, Guangdong, Yunnan, Heilongjiang and Hubei provinces were selected by multi-stage cluster random sampling method. Data were obtained on demographic information, neglect, physical abuse and non-suicidal self-injury from questionnaire survey. Results: During the past year before survey, 13.5% of participants engaged in self-injury 5 times or more, 15.4% of participants engaged in self-injury 1 to 4 times. Both neglect and physical abuse were significantly associated with self-injury in adolescents(P<0.01). After adjusted for gender, ethnicity, the only-one child, family structure, family economic status, and other variables, multinomial logistic regression showed that neglect and physical abuse were positively associated with self-injury behavior(OR=1.43-3.23). There was a potential dose-response relationship of accumulating physical abuse types and self-injury, with more physical abuse types, the higher risk of self-injury behavior. Conclusion Neglect and physical abuse experience should be considered as a risk factor for NSSI among adolescents in rural China. Specific intervention targeting neglect and physical abuse should be strengthened to reduce the occurrence of NSSI.

Objective: To investigate the effect and significance of GSK-3β inhibitor(LiCl)and RANK-RANKL on the renal tissue of diabetic nephropathy(DN) rats. Methods: SD rats were divided into normal control group (NC), DN model group (DN) and GSK-3β inhibitor intervention group (LiCl). Twenty-four hour urine protein of rats were determined by Coomassie brilliant blue. Kidney tissue sections were stained by HE. The expression of GSK-3β, RANK and RANKL protein were determined by immunohistochemistry staining. The mRNA of GSK-3β, RANK, RANKL was detected by RT-qPCR. Results: Compared with NC group[(14.72±3.37)g], the level of 24-hour urinary protein[(154.17±20.65)g] increased significantly in DN group; compared with DN Group, the level of 24-hour urinary protein [(107.22±31.15)g]decreased in LiCl group(P<0.05). Compared with NC group(2.10±0.60, 1.10±0.20, 1.21±0.20; 19.52±3.20, 1.80±1.10, 1.81±0.50), the pathological changes of renal tissues of DN group aggravated, the mRNA and expression of protein of GSK-3β, RANK and RANKL increased(9.10±2.15, 8.95±2.40, 9.90±2.60; 32.70±7.20, 19.20±4.32, 20.92±5.90); compared with DN group, the pathological changes of renal tissues of LiCl group alleviated, mRNA and the expression of protein of factors above declined(2.70±0.80, 2.32±0.65, 3.58±1.10; 22.35±3.25, 4.20±2.42, 5.90±2.36; P<0.05). Conclusion RANK and RANKL play an important role in the development of DN, LiCl influence Wnt and NF-κB signal pathway down-regulating RANK and RANKL to suspend development of diabetic nephropathy.

Objective To investigate the expression level of receptor-interacting protein serine threonine kinase (RIP) 3 in macrophages/monocytes activation in autoimmune hepatitis (AIH) and its regulation on inflammatory cytokines.Methods The degree of macrophage infiltration and the expression of RIP3 in liver tissues from patients with AIH or hepatic cysts by double-immunofluorescence.After 24 hours treated with different concentrations (0,1,3,6,10 μg/mL) of lipopolysaccharide (LPS),Necrostatin-1,the specific inhibitor of RIP3 signaling pathway,and 6-thioguanine,the active metabolite of azathioprine,the expression levels of RIP1 (RIP3 upstream signal molecule),RIP3 and mixed-lineage kinase domain-like protein (MLKL,RIP3 downstream substrate) in RAW264.7 macrophages were detected by Western blotting.The expression of macrophage-associated cytokine at mRNA level of each treatment group was determined by real time quantitative polymerase chain reaction (PCR).Student's t test and sum rank test were performed for statistical analysis.Spearman analysis was performed for the correlation analysis.Results Compared with hepatic cysts adjacent liver tissues,the infiltration of CD68 positive macrophages in liver tissue of AIH patients was significantly increased (4.75 ± 0.96 vs 28.86 ± 6.23),and the difference was statistically significant (t =7.80,P＜0.05),and the expression level of RIP3 also was significantly increased (15,11 to 22 vs 0,0 to 1),and the difference was statistically significant (Z=-2.66,P＜0.05).In vitro,compared with those of control group,the expression levels of RIP1,RIP3 and MLKL stimulated by LPS at 0,1,3,6,10 μg/mL were significantly increased,and the differences were statistically significant (t=4.00,4.90,6.40,10.30;3.80,9.30,9.80,9.00;4.90,9.90,9.30 and 7.70;all P＜0.05),and were dose-dependent (r=0.91,0.86 and 0.79,all P＜0.05).Furthermore,compared with those of LPS-stimulated group,the expressions of RIP1,RIP3,MLKL of LPS+Necrostatin-1 group were significantly decreased (0.73±0.11 vs 0.47±0.13,0.60±0.07 vs 0.37 ± 0.05,0.65 ± 0.22 vs 0.38 ± 0.04,respectively),and the differences were statistically significant (t=2.60,4.50 and 2.10,all P＜0.05).And the expressions of interleukin (IL)-1β,IL-6and IL-10 at mRNA levels were also decreased (810.3±200.8 vs 463.7±118.1,1 504.4±482.7 vs 290.4±106.9,1 358.6 ± 559.2 vs 677.8 ± 297.6,respectively),and the differences were statistically significant (t=5.40,12.52,5.70,all P＜0.05).However,the expressions of IL-4 and TGF-β at mRNA levels up-regulated (0.3±0.2 vs 0.6±0.3,0.4±0.1 vs 0.9±0.4,respectively),and the differences were statistically significant (t=4.60 and 6.10,both P＜0.05).Compared with those of the LPS-stimulated group,the expressions of IL-1β,IL-6 and IL-10 at mRNA levels of LPS and 6-thiopurine stimulated group significantly down-regulated (810.3±200.8 vs 283.4±65.5,1 504.4±482.7 vs 354.4±73.8,1 358.6± 559.2 vs 625.6±336.3),and the differences were statistically significant (t=4.30,10.60 and 3.50,all P＜0.05);however,the expressions of IL-4 and TGF-β at mRNA levels significantly up-regulated (0.3±0.2 vs 0.6±0.1 and 0.4±0.1 vs 0.5±0.1),and the differences were statistically significant (t=5.20and 12.50,P＜0.05).Conclusions The regulation effects of 6-thiopurine on RIP3 signaling pathway and related cytokines are similar to those of Necrostatin-1.And the expression of RIP3 signaling protein increasing in activated macrophages of liver tissues from AIH patients is closely related to the regulation of IL-6.The RIP3 mediated inflammatory signaling pathway in macrophage may be involved in the genesis and development of AIH and may be a potential therapeutic target.