Objective To investigate the association of serum leptin (LEP) levels during the first postpartum year with the occurrence of postpartum thyroiditis (PPT).Methods Fifty-seven PPT patients consisted of 34 with overt PPT and 23 subclinical PPT.37 healthy postpartum women were used as controls.Serum samples were obtained at 4 postpartum date points,i.e.3-days and 3,6,12-months postpartum.LEP level was determined by radioimmunoassav.Results Compared with control women,PPT patients were maintaining significantly higher levels of LEP and LEP/body mass index (BMI) ratio during the first postpartum year.There was no significant difference in serum LEP level or LEP/BMI ratio between overt PPT and subclinical PPT groups.In PPT patients,LEP and LEP/BMI ratio were negatively correlated with serum TSH,and positively correlated with serum FT4 and FT3.Conclusion Sustained high levels of serum LEP after delivery may favor the occurrence of PPT.Further studies are needed to clarify the specific role played by LEP in PPT.

Objective To investigate the relationship between genetic variants in the Toll-like receptor (TLR) pathway genes and susceptibility of gastric cancer (GC) and esophageal squamous cell carcinoma (ESCC).Methods The data of whole genome association studies of the high-risk population of GC and ESCC in China were analyzed by adaptive rank-truncated product (ARTP) method in pathway and gene level.The associations between single nucleotide polymorphism (SNP) and susceptibility of GC and ESCC were analyzed with additive model of unconditional Logistic regressions.PLINK 1.07 and SPSS 19.0 software were performed for statistical analyses,and ARTP package in R3.0.2 was used for pathway and gene level analysis.Results In gene-level analyses,eight genes were found to be associated with susceptibility of GC (P ＜0.05) and six genes were associated with susceptibility of ESCC (P ＜ 0.05).In single SNP-level analyses,21 SNPs were statistically correlated with susceptibility of GC (P ＜ 0.01),and 11 SNPs were statistically correlated with susceptibility of ESCC (P ＜0.01).Conclusions Some genetic variants in TLR pathway are associated with risk of GC and ESCC.The potential molecular mechanisms need further investigation.

ObjectiveTo investigate the cumulative incidence of persistent hypothyroidism in patients who were diagnosed as postpartum thyroiditis ( PPT),and to determine the factors associated with the development of persistent hypothyroidism in those patients.MethodsThe present study was performed as the continuous study followed by the former epidemiological survey on PPT,in which 58 patients with PPT (35 overt PPT and 23 subclinical PPT) were diagnosed.The 58 patients were followed up at 12th month postpartum,and then for every 6 months until 24 months postpartum.Fasting blood samples were taken for testing serum TSH,thyroid peroxidase antibody ( TPOAb),and thyroglobulin antibody ( TgAb ).Free T3 ( FT3 ),free T4 ( FT4 ),and TSH receptor antibody ( TRAh ) were detected if TSH was abnormal.50 healthy postpartum women were used as control group.ResultsOf the total 58 PPT patients,91,4％ ( n =53 ) were successfully followed.Five patients with overt PPT and 6 patients with subclinical PPT developed persistent hypothyroidism,and the cumulative incidence of persistent hypothyroidism in the studied PPT patients was 20.8％.Among 15 PPT patients who had a classical biphasic course (a thyrotoxic phase followed by a hypothyroid phase),persistent hypothyroidism was seen in 26.7％ (n =4 ).Among 11 PPT patients with hypothyroidism only,persistent hypothyroidism was seen in 63.6％ ( n =7).On the contrary,none of the patients with thyrotoxicosis only had persistent hypothyroidism.All of the patients who developed persistent hypothyrodism had a higher TSH levels than 4.8 mU/L at 6th month postpartum.Before delivery,TSH levels of the patients developed persistent hypothyroidism were significantly higher than those of the patients with transient hypothyroidism,and this was the case at the 12th month postpartum ( all P＜0.01 ).PPT patients maintained a relatively higher rate of thyroid autoantibodies.The positive rate of TPOAb at the 12th,18th,and 24th month postpartum was 56.6％,50.9％,and 52.8％,respectively; and the positive rate of TgAb being 35.8％,30.2％,and 30.2％,respectively.Both the positive rate and titer of TPOAb in patients with overt PPT were higher than those in patients with subclinical PPT at the 18th and 24th month postpartum (P＜0.05).Conclusions 20.8％ patients with PPT developed persistent hypothyroidism at the 24th month postpartum.Whether a patient with PPT would develop persistent hypothyroidism depends on his clinical feature and TSH level.

Objective: To study the mechanism of interleukin (IL)-17 in mice with non-alcoholic fatty liver disease for promoting M1-type macrophage polarization to exacerbate liver inflammation, and to provide references for the mechanism of NAFLD occurrence and development. Methods: A mouse model of NAFLD was constructed by high-fat diet. Mice were divided into control group, model group, IL-17 group, and anti IL-17 group. Histopathological changes of the liver were observed by HE staining. The serum levels of ALT and AST in peripheral blood of mice was detected by chemical colorimetry. Macrophages labeled with F4/80-PE, CD11C-FITC was designated as M1-type macrophages, those labeled with F4/80-PE, and CD206-APC was designated as M2-type macrophages. The proportion of M1 and M2 macrophages infiltrated into the liver tissues of mice were measured by flow cytometry. CD168 expression level of liver tissues was detected using immunohistochemistry. Protein and mRNA levels of the marker molecules (iNOS, TNF-alpha and IL-6) of M1 macrophages were detected using ELISA and RT-Q PCR. Western blot was used to detect the protein expression of JAK-STAT signal pathway and the expression level of MCP-1. Data were analyzed using one-way ANOVA and t-test. Results: High-fat diet NAFLD mice model was successfully constructed. IL-17 had increased the proportion of M1 macrophages in mice liver tissues and decreased the proportion of M2 macrophages (P < 0.05). The proportion of M1 and M2 macrophages in the liver tissues of normal mice was 7.9% ± 1.1% and 19.2% ± 1.8%. The proportion of M1 and M2 macrophages in the model group was 17.3% ± 2.5% and 15.0% ± 2.1. The proportion of M1 macrophages (33.8% ± 4.2%) in IL-17 group was higher than model group, while the proportion of M2 macrophages (7.8% + 1.0%) in IL-17 group was lower than model group. Protein and mRNA marker levels of M1 macrophage (iNOS, IL-12, TNFα and IL-6) in liver tissues were significantly higher than model group, control group, and anti-IL-17 group (P < 0.05). The expression levels of JAK1, STAT1, MCP-1, and CD168 in mice liver tissues of IL-17 group had increased (P < 0.05). The levels of aspartate and alanine aminotransferases in peripheral blood of mice in IL-17 group were significantly higher than other three groups (P < 0.05). Conclusion IL-17 can promote M1-type macrophage polarization, and exacerbates the liver inflammatory response to accelerate the progression of NAFLD in mice.

OBJECTIVES: Pseudolaric acid B (PAB) has been shown to inhibit the growth of various tumor cells, but the molecular details of its function are still unknown. This study investigated the molecular mechanisms by which PAB induces apoptosis in HeLa cells. METHODS: The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assays were performed to investigate the effect of PAB treatment in various cervical cancer cell lines. Annexin V/propidium iodide staining combined with flow cytometry and Hoechst 33258 staining were used to assess PAB-induced apoptosis. Additionally, we performed bioinformatics analyses and identified a paired box 2 (PAX2) binding site on the BAX promoter. We then validated the binding using luciferase and chromatin immunoprecipitation assays. Finally, western blotting assays were used to investigate PAB effect on the Wnt signaling and the involved signaling molecules. RESULTS: PAB promotes apoptosis and downregulates PAX2 expression in HeLa cells in a time- and concentration-dependent manner. PAX2 binds to the promoter of BAX and inhibits its expression; therefore, PAX2 inhibition is associated with increased levels of BAX, which induces apoptosis of HeLa cells via the mitochondrial pathway. Additionally, PAB inhibits classical Wnt signaling. CONCLUSION: PAB effectively inhibits Wnt signaling and PAX2 expression, and increases BAX levels, which induce apoptosis in HeLa cells. Therefore, PAB is a promising natural molecule for the treatment of cervical cancer.
Apoptosis ; Binding Sites ; Bisbenzimidazole ; Blotting, Western ; Cell Line ; Chromatin Immunoprecipitation ; Computational Biology ; Flow Cytometry ; HeLa Cells ; Humans ; Luciferases ; Mitochondria ; Uterine Cervical Neoplasms ; Wnt Signaling Pathway

Objective To study the mechanism of minocycline inhibiting inflammatory reaction of trigeminal ganglion glial cells in trigeminal neuralgia rats,and provide reference and support for treatment of trigeminal neuralgia.Methods (1) The rat models of trigeminal neuralgia were established by laser chemical induction oftrigeminal nerve injury.Thirty SD rats were randomly divided into sham-operated group,model Ⅰ group,minocycline group (intragastric administration of 50 μg minocycline for 7 d,n=10).The threshold of mechanical pain was measured in the facial nerve areas of rats.The protein and mRNA expressions of nuclear transcription factor (NF)-κB and interleukin (IL)-1β in the trigeminal ganglion were detected,the activation of satellite glial cells was observed by glial fibrillary acidic protein (GFAP) staining,and immunohistochemical staining was used to detect NF-κB level.(2) The inflammatory models of glial cells were established with nitroglycerin;and trigeminal glial cells from the SD rats were cultured in vitro;the cell models were divided into control group,model Ⅱ group,low-dose minocycline group (15 μmol/L),and high-dose minocycline group (30 μmol/L);the expressions of NF-κB and IL-1β in cells were detected.Fluo-3/AM probe load was used to observe the concentration changes of calcium ions in the glial cells.Results (1) The threshold of pain in trigeminal neuralgia of minocycline group was significantly higher than that of model Ⅰ group (P＜0.05);the protein and mRNA expressions of NF-κB and IL-1β in the minocycline group were significantly decreased as compared with those in model Ⅰ group (P＜0.05);weak NF-rB expression was noted in the sham-operated group,strong NF-κB expression was noted in the model Ⅰ group,and that in the minocycline group was obviously decreased as compared with that in model Ⅰ group;number of GFAP positive cells in the minocycline group was significantly smaller as compared with the model Ⅰ group (P＜0.05).(2)The protein and mRNA expressions of NF-κB and IL-1β in the low-dose minocycline group and high-dose minocycline group were significantly decreased as compared with those in the model Ⅱ group (P＜0.05);and the concentration of calcium ions in astrocytes of the low-dose minocycline group and high-dose minocycline group was significantly decreased as compared with that of model Ⅱ group (P＜0.05).Conclusion Minocycline can alleviate pain in trigeminal neuralgia rats by inhibiting the activation of satellite glial cells and decreasing the levels of inflammatory factors NF-κB and IL-1β.

Objective To study the effect of intestinal bacteria on motor ability of amyotrophic lateral sclerosis (ALS) mice models and its mechanism. Methods Twenty wild type C57BL/6J mice (WT group) and 20 SOD1-G93A transgenic ALS mice (ALS group) were selected as the research subjects. (1) Ten mice in both WT group and ALS group were selected, respectively; 5 mice in each group were fed in SPF environment, and the remaining 5 mice were fed in aseptic environment; they were defined as WT+SPF group, WT+aseptic group, ALS+SPF group and ALS+aseptic group. (2) Ten mice in WT group and ALS group were fed in sterile environment; 5 mice in each group were transplanted with fecal bacteria, and the remaining 5 mice were not interfered; they were defined as WT+transplantation group, WT+non-transplantation group, ALS+transplantation group and ALS+non transplantation group. The grip strength of mice was measured by grip force meter, the motor coordination ability of mice was tested by roller treadmill and rotating rod test, the number of motor neurons in the anterior horn of spinal cord was measured by Nissl staining, the expression of microglia activation marker ionic calcium junction protein (IBA-1) in spinal cord tissues was detected by immunohistochemical staining, and the expressions of tumor necrosis factor (TNF)-α and interleukin (IL)-6 in spinal cord tissues were detected by Western blotting; the β-N-methylamino-L-alanine (BMAA) expression was detected by high performance liquid chromatography-tandem mass spectrometry. Results (1) The grip strength, drop latency and drop time of ALS+aseptic mice were significantly higher than those of ALS+SPF mice, the number of Nissl-stained positive cells was significantly larger than that of ALS+SPF mice, the number of IBA-1 positive cells was significantly smaller than that of ALS+SPF mice, the levels of TNF-α and IL-6 protein expressions and BMAA concentration were statistically lower than those of ALS+SPF mice (P<0.05). (2) The grip strength, drop latency and drop time of ALS+transplantion mice were significantly lower than those of ALS+non-transplantation mice, the number of Nissl-stained positive cells was significantly smaller than that of ALS+non-transplantation mice, the number of IBA-1 positive cells was significantly larger than that of ALS+non-transplantation mice, the TNF-α and IL-6 protein expressions and BMAA concentration were significantly higher than those of ALS+non-transplantation mice (P<0.05). Conclusion Imbalance of intestinal bacteria homeostasis can decrease the motor ability of ALS mice, which is related to the activation of microglia.

Objective To study the correlation of the prognosis of Icotinib administration with the expression levels of F-box and WD repeat domain-containing 7(FBW7) and myeloid cell leukemia-1 (MCL-1) in peripheral blood in elderly patients with advanced non-small-cell lung cancer.Methods A total of 76 patients aged 60 years or over diagnosed with non-small-cell lung cancer(NSCLC) with EGFR-sensitive mutations and under Icotinib treatment were enrolled in this study.FBW7 and MCL-1 mRNA expression levels in peripheral blood were detected by real-time quantitative PCR(RT-QPCR).The correlation of FBW7 and MCL-1 expression levels with clinical and histological parameters,overall survival (OS),and progression-free-survival (PFS) was analyzed.Results The FBW7 expression level and the MCL-1 expression level were negative correlated(r =-0.37,P ＜0.001).High FBW7 expression levels and low MCL-1 expression levels in peripheral blood were associated with improved therapeutic efficacy of Icotinib (P＜0.001) and extended OS and PFS.Cox regression analysis showed that the expression levels of FBW7 and MCL-1 in peripheral blood were independent influencing factors for OS and PFS.Conclusions Patients with high FBW7 expression levels and low MCl-1 expression levels are more likely to benefit from Icotinib treatment.Expression levels for either factor can be used as a predictive indicator for the effectiveness of Icotinib and provide guidance for its clinical use.

Objective To investigate the proportion of helper T cell subset Th 9 and the expression of its cytokine interleukin-9(IL-9)in Parkinson's disease(PD)and the clinical significance.Methods Seventy-two patients diagnosed with PD between January 2016 and June 2017 and 20 healthy volunteers in the same period were selected.The PD patients were staged according to the Hoehn-Yahr(H-Y)staging method,21 in stage one,19 in stage two,18 in stage three,11 in stage four,and three in stage five.The proportion of Th9 subset in peripheral blood of PD patients and healthy volunteers was measured by flow cytometry.The expression of IL-9 in peripheral blood of PD patients and healthy volunteers was measured by enzyme-linked immunosorbent assay.Results The proportion of Th9 cells in peripheral blood of PD patients(1.27%±0.34%)was significantly higher than that of healthy volunteers(0.61%±0.11%,t=8.530,P<0.05),and the higher stage of PD,the higher proportion of Th9,suggesting that the proportion of Th9 was related to the PD staging.IL-9 was also highly expressed in PD patients((16.04 ±2.94) pg/ml)and had statistically significant difference compared with healthy volunteers((7.53 ±0.70)pg/ml;t=12.781,P<0.05).IL-9 was similar to Th9, the higher stage of PD, the higher expression of IL-9. Conclusion The proportion of Th9 cells and the expression of IL-9 in peripheral blood of patients with PD increased significantly,having a significant relationship with the H-Y staging of PD.

Objective To investigate the effects of dl-3-n-butylphthalide ( NBP) on angiogenesis of human umbilical vein endothelial cells ( HUVEC) in vitro under ischemia/hypoxia and the correlation with vascular endothelial growth factor/vascular endothelial growth factor receptor 2 ( VEGF/VEGFR2 )-Notch1/Dll4 signaling pathway .Methods The control group , ischemia/hypoxia group , and ischemia/hypoxia +NBP group ( high dose group and low dose group ) were set up after HUVEC subculture . The cell concentration was adjusted to be 1 ×105/ml, and each group was randomly added 1 ml (1 ×105 cells in each group).Both of the ischemia/hypoxia group and the ischemia/hypoxia +NBP group were cultured under the condition of ischemia and hypoxia .The NBP concentrations of the high and low dose groups were 20 μmol/L and 5 μmol/L respectively.The cell viability of each group was detected by cell counting kit-8, and cell scratch test was used to detect the migration ability of the cells in each group .The cell formation ability of each group was examined by in vitro angiogenesis assay.Western blotting was used to detect the expressions of receptor proteins VEGFR2, Notch1 and Dll4,and mRNA expressions of VEGF, VEGFR2, Notch1 and Dll4 were detected by real-time quantitative PCR.Results NBP increased the survival rates of HUVEC under ischemia/hypoxia condition.In the low dose group, the survival rates of HUVEC at 6, 12, 24, 48 h were 78.6％ ±3.0％, 59.6％ ±5.3％, 44.6％ ±4.2％, 38.2％ ±4.3％, respectively, significantly higher than that in the ischemia/hypoxia group (75.2％±5.8％, 53.2％±4.8％, 36.2％± 7.8％, 22.5％±4.1％;t=4.513, 6.231, 9.322, 9.674; P=0.021, 0.018, 0.026, 0.015).In the high dose group, the survival rates of HUVEC at 6, 12, 24, 48 h were 88.6％±6.3％, 67.5％±5.4％, 53.3％±4.2％, 46.3％±3.9％, respectively , also significantly higher than that in the ischemia/hypoxia group (t=8.123, 11.211, 12.312, 14.154;P=0.001, 0.002, 0.001, 0.001).The mobility of the low dose group was 52.3％+4.2％, with statistically significant difference compared with that in the ischemia /hypoxia group ( 18.5％ ±3.2％) and the control group ( 22.3％ ±4.1％; t=18.324, 15.183; P=0.000, 0.000).The mobility of the high dose group was 87.5％±5.2％, also with statistically significant difference compared with that in the ischemia/hypoxia group and the control group ( t=22.142, 19.341;P=0.000, 0.000).NBP increased the protein and mRNA expression of VEGF , VEGFR2, Notch1 and Dll4.The relative expression of VEGFR2, Notch1 and Dll4 in the low dose group was 1.12 ±0.17, 0.35 ± 0.07 and 0.42 ±0.08, respectively, with statistically significant difference compared with that in the ischemia/hypoxia group (0.82 ±0.05, 0.30 ±0.03, 0.32 ±0.04;t=6.120, 2.123, 4.112;P=0.000, 0.020, 0.003).The relative expression of VEGFR2, Notch1 and Dll4 in the high dose group was 1.30 ± 0.15, 0.41 ±0.10 and 0.48 ±0.11, respectively, also with statistically significant difference compared with that in the ischemia/hypoxia group ( t=8.122, 3.851, 5.130; P=0.000, 0.000, 0.001 ) .The mRNA expressions of VEGF , VEGFR2, Notch1 and Dll4 in the low dose group were 0.43 ±0.08, 0.41 ± 0.05, 0.38 ±0.03 and 0.36 ±0.04, respectively, with statistically significant difference compared with that in the ischemia/hypoxia group (0.28 ±0.03, 0.34 ±0.04, 0.27 ±0.03, 0.19 ±0.04;t=3.122, 3.825, 4.311, 5.211; P=0.000, 0.006, 0.001, 0.000).And the mRNA expressions of VEGF, VEGFR2, Notch1 and Dll4 in the high dose group were 0.58 ±0.05, 0.50 ±0.06, 0.41 ±0.05, 0.52 ±0.06, respectively, also with statistically significant difference compared with that in the ischemia /hypoxia group (t=4.225, 4.872, 5.311, 8.220;P=0.000, 0.000, 0.000, 0.000).Conclusions NBP can promote HUVEC to form blood vessels under ischemia/hypoxia condition , the mechanism of which may be related to the activation of VEGF/VEGFR2-Notch1/Dll4 signaling pathway .It may be one of the mechanisms that NBP improves cerebral microcirculation in acute ischemic stroke .