Comparative assessment of gut microbial composition and function in patients with Graves' disease and Graves' orbitopathy.

BACKGROUND: A previous study indicated that gut microbiota changed notably in Graves' orbitopathy (GO) patients as compared to controls. However, the characteristics of intestinal bacteria in Graves' disease (GD) and GO are unclear. OBJECTIVE: The present study aimed to identify specific intestinal bacteria of GD and GO, respectively. METHODS: The gut microbial communities of the fecal samples of 30 GD patients without GO, 33 GO subjects, and 32 healthy subjects were analyzed and compared by 16S rRNA gene sequencing. RESULTS: At the phylum level, the proportion of Deinococcus-Thermus and Chloroflexi was decreased significantly in GO patients as compared to GD. At the genus level, the proportion of Subdoligranulum and Bilophila was increased while that of Blautia, Anaerostipes, Dorea, Butyricicoccus, Romboutsia, Fusicatenibacter, unidentified_ Lachnospiraceae, unidentified_Clostridiales, Collineslla, Intestinibacter, and Phascolarctobacterium was decreased in the GO group as compared to the GD group. Random forest analysis was used for the identification of specific intestinal microbiota, and Deinococcus-Thermus, Cyanobacteria and Chloroflexi were ranked in the top ten according to their contributions to sample classification. Moreover, compared to the control, there were multiple gut bacterial enrichment metabolic pathways in GO and GD patients, including nucleotide metabolism, enzyme family, and energy metabolism. Compared to GO, the only enrichment metabolic pathway found in GD was the viral protein family. CONCLUSIONS: This study highlighted the significant differences in the intestinal microbiota and predictive functions of GD with GO, thereby providing new insights into the role of the gut bacteria that might contribute to the development of GO in GD patients.

[Changes and clinical significance of peripheral blood CD8(+)CD25(+)T cells in rheumatoid arthritis patients].

Objective: To investigate the expression of CD8(+)CD25(+)T cells in peripheral blood of patients with rheumatoid arthritis (RA) and its correlation with clinical indicators of rheumatoid arthritis. Methods: Peripheral blood was collected from 38 patients with RA, and 20 healthy control subjects, RA patients admitted to Peking University people's hospital from May to October 2018, and record the RA patients with the clinical manifestations and laboratory indexes, extraction in the peripheral blood lymphocytes, using flow cytometry to analyse the percentage of CD8(+)CD25(+)T cells in peripheral blood, by using the software SPASS20 and Prism6 to analyze its correlation with clinical and laboratory indices. Results: The expression of CD8(+)CD25(+)T cells in peripheral blood of RA patients was significantly increased, which was statistically different from that of healthy patients (P<0.05). CD8(+)CD25(+)T cells in peripheral blood of RA patients showed significant positive correlation with ESR(r=0.352,P=0.030), CCP(r=0.312,P=0.047) and DAS28(r=0.330,P=0.043), and negatively correlated with C3 (r=-0.354,P=0.046) and C4(r=-0.440,P=0.010).No significant correlation was found in other indicators. In RA patients, there were statistically significant differences in CD8(+)CD25(+)T cells between the low-disease active group and the high-disease active group(P<0.05), but CD8(+)CD25(+)T cells between the low-disease active group and the moderate-disease active group, or between the moderate-disease active group and the high-disease active group had no significant statistical difference. Conclusion: CD8(+)CD25(+)Tcells in peripheral blood of patients with RA are significantlyincreased, and aresignificantly correlated with laboratory and clinical indicators, which may play an important role in the pathogenesis of rheumatoid arthritis.

Alterations in the intestinal microbiota of patients with severe and active Graves' orbitopathy: a cross-sectional study.

BACKGROUND: The intestinal microbiota was linked to autoimmune diseases. Graves' orbitopathy (GO) is an autoimmune disease that is usually associated with Graves' disease. However, information on the microbiome of GO patients is yet lacking. OBJECTIVES: To investigate whether GO patients differ from healthy controls in the fecal microbiota. DESIGN: A cross-sectional study. SETTING: 33 patients with severe and active GO and 32 healthy controls of Han nationality were enrolled between March 2017 and March 2018. METHODS: The Gut microbial communities of the fecal samples of GO patients and healthy controls were analyzed and compared by 16S rRNA gene sequencing. RESULTS: Community diversity (Simpson and Shannon) was significantly reduced in fecal samples from patients with GO as compared to controls (p < 0.05). The similarity observed while assessing the community diversity (PCoA) proposed that the microbiota of patients with GO differ significantly from those of controls (p < 0.05). At the phyla levels, the proportion of Bacteroidetes increased significantly in patients with GO (p < 0.05), while at the genus and species levels, significant differences were observed in the bacterial profiles between the two groups (p < 0.05). LIMITATIONS: Single-centered study design and limited fecal samples. CONCLUSIONS: The present study indicated distinctive features of the gut microbiota in GO patients. The study provided evidence for further exploration in the field of intestinal microbiota with respect to the diagnosis and treatment of GO patients by modifying the microbiota profile.

Effect of night light regimen on growth performance, antioxidant status and health of broiler chickens from 1 to 21 days of age.

OBJECTIVE: The study was conducted to evaluate the effects of night light regimen on growth performance, antioxidant status and health of Lingnan Yellow broiler chickens from 1 to 21 days of age. METHODS: A completely randomized factorial design involved 2 photoperiods (constant lighting [CL], 24 L:0 D and intermittent lighting [INL], 17 L:3 D:1 L:3 D)×2 light intensities (10 lx and 30 lx). A total of one thousand six hundred and eighty 1-d-old Lingnan Yellow broiler chicks were randomly divided into 4 treatments with 6 replicates (70 birds per replicate). The experiment lasted for 21 d. RESULTS: Photoperiods and light intensities had no effect on average daily gain, feed conversion ratio, and mortality of the broiler chickens (p>0.05). The INL had a significant effect on average daily feed intake (p<0.05) of broiler chickens compared with CL. Photoperiod and light intensity had an interactive effect on melatonin (MT) concentration (p<0.05). At CL, reducing light intensity increased MT concentration; INL birds had higher MT but MT concentration was not affected by light intensity. There was an interactive effect on glutathione peroxidase (GPx) and catalase (CAT) in serum and total antioxidant capability (T-AOC) in liver between photoperiod and light intensity. With the decrease of light intensity, the activities of GPx and CAT in serum and T-AOC in liver increased in CL group (p<0.05). Broiler chickens reared under INL had better antioxidant status and 10 lx treatments had higher activities of CAT in serum than 30 lx (p<0.05). Different photoperiods and light intensities had no effect on malondialdehyde. There was an interaction between photoperiod and light intensity on serum creatine kinase (CK) concentration (p<0.05). At CL, the elevated light intensity resulted in an increase in CK content; INL birds had lower CK concentration especially in low light intensity group. Besides, INL and low light intensity significantly reduced the concentration of serum corticosterone and heat shock protein 70 (p<0.05). Serum immunoglobulin M contents were increased in broiler chickens reared under the INL compared with CL group (p<0.05). CONCLUSION: Results above suggest that the night light regimen of INL and 10 lx could be beneficial to the broiler chickens from 1 to 21 days of age due to the better health status and electricity savings.

Effects of different selenium sources and levels on antioxidant status in broiler breeders.

OBJECTIVE: This study was conducted with the objectives to examine the impacts of inorganic selenium (Se) and different types and levels of organic selenium on the serum and tissues Se status and antioxidant capacity in broiler breeders. METHODS: Five hundred and forty 48-wk-old Lingnan Yellow broiler breeders were randomly assigned to 6 dietary treatments, provided same basal diet (0.04 mg/kg of Se) with 0.15 mg/kg, or 0.30 mg/kg of Se from sodium selenite (SS) or from selenium-enriched yeast (SY) or from selenomethionine (SM). The broiler breeders were slaughtered after an 8-wk experiment. RESULTS: The results showed that SM was better than SY and SS, 0.30 mg/kg level was better than 0.15 mg/kg level in Se deposition (p<0.05) in serum, liver, kidney, pancreas and muscle; in antioxidant status, organic selenium had better effects than SS in broiler breeders (p<0.05), but SM and SY had a similar result, and 0.15 mg/kg level was better than 0.30 mg/kg (p<0.05). CONCLUSION: The results demonstrated the evident advantage of supplementation of broiler breeders with 0.15 mg/kg SM, which improved tissue Se concentrations and antioxidant status, and can be considered as the best selenium source.

The effects of the polymorphism in exon 3 of the FAS gene on the death of chicken embryos during the incubation period.

To investigate the effect of a factor-associated suicide (FAS) gene polymorphism on the death of chicken embryos, we genotyped 190 dead embryos and 69 normally developing embryos from 7200 hatching Short-Leg Yellow Chicken eggs, as well as 119 dead embryos and 69 normally developing embryos from 4650 hatching Yellow B Chicken eggs. The results showed that there were significant (P < 0.05) genotypic differences between dead and normally developing embryos for this FAS gene polymorphism, a SNP in exon 3 (NC_006093.2:g.6514A>C, rs15793179). Logistic regression revealed that Short-Leg Yellow Chicken embryos with genotype g.6514CC had a significantly (P < 0.01) higher risk of death than those with genotype g.6514AC. This polymorphism has the potential to be an effective tool when used in conjunction with traditional selection methods.

Sex ratio bias in early-dead embryos of chickens collected during the first week of incubation.

According to Mendelian heredity laws, the sex ratio of a given chicken population during hatching is expected to be 1:1. In this study, we collected 432 chicken embryos that died during the first week of incubation from 5 different breeds. The sexes of the early-dead embryos were determined by using the previously described molecular sexing technique of double PCR. The female-to-male sex ratio was analyzed for departure from the expected 1:1 sex ratio by chi(2) testing. These results showed that the number of female dead embryos was significantly greater than that of males in the Hubei local breeding stock, Zhusi, and Hy-line Variety Brown (P < 0.05, P < 0.01, P < 0.01 respectively), with observed female-to-male sex ratios of 1.40:1, 2.03:1, and 2.22:1, respectively. Two other Chinese local breeds (the Yellow chicken and the Aijiaohuang chicken) also showed altered sex ratios, although the differences were not significant. Altogether, these results indicated that female chickens were more likely than male chickens to die at the early stages of incubation.

CDH17 is a downstream effector of HOXA13 in modulating the Wnt/ß-catenin signaling pathway in gastric cancer.

OBJECTIVE: In this study, we investigated the mechanism underlying co-upregulation of HOXA13 and CDH17 in gastric cancer, the signaling pathway in which HOXA13 and CDH17 involve in and their functional role in gastric cancer cells. MATERIALS AND METHODS: Relevant microarrays investigated the dysregulated genes in gastric cancer tissues were searched in ArrayExpress. The co-expression of HOXA13 and CDH17 was analyzed in the gastric cancer patient cohort in TCGA database using cBioportal and UCSC Xena. The regulative effect of HOXA13 on CDH17 expression was examined by dual luciferase assay. The involvement of HOXA13 and CDH17 in the Wnt/beta-catenin signaling pathway was assessed by Western blotting. The functional role of HOXA13 and CDH17 in gastric cancer cells were studied by CCK-8 assay of cell growth, Transwell assay of cell invasion and flow cytometry of active caspase-3. RESULTS: HOXA13 and CDH17 expression are upregulated and are highly correlated in gastric cancer tissues. HOXA13 overexpression significantly increased CDH17 mRNA and protein expression and also significantly increased the transcription activity of the luciferase reporter with integrate HOXA13 binding sites. HOXA13 shRNA and CDH17 shRNA had similar effect on reducing the expression of beta-catenin, while shCDH17 abrogated HOXA13 induced upregulation of beta-catenin. HOXA13 shRNA and CDH17 shRNA decreased cell proliferation and invasion and increased cell apoptosis in SGC-7901 cells. CONCLUSIONS: HOXA13 can elevate CDH17 transcription via binding to its promoter. CDH17 is a downstream effector of HOXA13 in modulating the Wnt/beta-catenin signaling pathway in gastric cancer cells. Both HOXA13 shRNA and CDH17 shRNA can decrease gastric cancer cell proliferation and invasion and increase their apoptosis.

[A review of progress in research of effect of air pollutants on fetal growth].

Slow fetal growth is a global public health concern because it might be associated with many diseases later in life. With the progress of technology to measure long-term air pollution exposure. Many epidemiological studies have evaluated the effects of maternal air pollutant exposure on fetal growth. In this paper, we summarize the research progress in this field after the analysis on the related literatures retrieved from the databases of Medline and Web of Science.

LKB1 inhibits HPV-associated cancer progression by targeting cellular metabolism.

Liver kinase B1 (LKB1) is mutationally inactivated in Peutz-Jeghers syndrome and in a variety of cancers including human papillomavirus (HPV)-caused cervical cancer. However, the significance of LKB1 mutations in cervical cancer initiation and progress has not been examined. Herein, we demonstrated that, in mouse embryonic fibroblasts, loss of LKB1 and transduction of HPV16 E6/E7 had an additive effect on constraining cell senescence while promoting cell proliferation and increasing glucose consumption, lactate production and ATP generation. Knockdown of LKB1 increased and ectopic expression of LKB1 decreased glycolysis, anchorage-independent cell growth, and cell migration and invasion in HPV-transformed cells. In the tumorigenesis and lung metastasis model in syngeneic mice, depletion of LKB1 markedly increased tumor metastatic colonies in lungs without affecting subcutaneous tumor growth. We showed that HPV16 E6/E7 enhanced the expression of hexokinase-ll (HK-II) in the glycolytic pathway through elevated c-MYC. Ectopic LKB1 reduced HK-II along with glycolysis. The inverse relationship between HK-II and LKB1 was also observed in normal and HPV-associated cervical lesions. We propose that LKB1 acts as a safeguard against HPV-stimulated aerobic glycolysis and tumor progression. These findings may eventually aid in the development of therapeutic strategy for HPV-associated malignancies by targeting cell metabolism.

LKB1 reduces ROS-mediated cell damage via activation of p38.

Liver kinase B1 (LKB1, also known as serine/threonine kinase 11, STK11) is a tumor suppressor mutated in Peutz-Jeghers syndrome and in a variety of sporadic cancers. Herein, we demonstrate that LKB1 controls the levels of intracellular reactive oxygen species (ROS) and protects the genome from oxidative damage. Cells lacking LKB1 exhibit markedly increased intracellular ROS levels, excessive oxidation of DNA, increased mutation rates and accumulation of DNA damage, which are effectively prevented by ectopic expression of LKB1 and by incubation with antioxidant N-acetylcysteine. The role of LKB1 in suppressing ROS is independent of AMP-activated protein kinase, a canonical substrate of LKB1. Instead, under the elevated ROS, LKB1 binds to and maintains the activity of the cdc42-PAK1 (p21-activated kinase 1) complex, which triggers the activation of p38 and its downstream signaling targets, such as ATF-2, thereby enhancing the activity of superoxide dismutase-2 and catalase, two antioxidant enzymes that protect the cells from ROS accumulation, DNA damage and loss of viability. Our results provide a new paradigm for a non-canonical tumor suppressor function of LKB1 and highlight the importance of targeting ROS signaling as a potential therapeutic strategy for cancer cells lacking LKB1.

Vocal fold epithelial hyperplasia. Vibratory behavior vs extent of lesion.

The vibratory behavior of 72 vocal folds with epithelial hyperplasia or dysplasia was investigated by means of videostroboscopy. The amplitude of vibration (AMP) and mucosal wave (WAV) were related to the relative area, depth, and relative volume of the lesion. The AMP and WAV were evaluated for the entire vocal fold (AMPE, WAVE) and for the affected portion or the lesion (AMPL, WAVL). The AMPE, AMPL, WAVE, and WAVL were negatively related to the relative area, depth, and relative volume. The relationship between the limited vibratory movement and the extent of the lesion was most significantly manifested in WAVL. A complete absence of any vibratory movement of the vocal fold took place only for large lesions occupying three fourths or more of the membranous vocal fold.

[Correlation of clinical findings and histologic types in epithelial hyperplasia of the vocal fold].

A series of 60 cases (85 vocal folds) with epithelial hyperplasia of the vocal fold were statistically analysed between the clinical findings and histologic types to identify the significant factors predicting the presence or absence of atypia in the lesion. The factors of age, sex, appearance of the lesion, extent of the lesion and degree of limitation of vocal fold vibration showed significance and therefore were scored respectively for further analysis from -1 to +1 according to the possibility of atypia from low to high. More than half of the group without atypia had total score of 0 or less, while more than half of the group with atypia had total score of +4 or more. The difference between two groups was significantly high (p less than 0.01). This results showed that if total score was 0 or less the possibility of atypia was very low and conservative treatment including discontinuation of smoking and follow-up can be conducted, whereas if total score was +4 or more the possibility of atypia was very high and a proper biopsy followed by laser surgery is indicated.

Liver kinase B1 regulates the centrosome via PLK1.

Liver kinase B1 (LKB1) is a tumor suppressor mutationally inactivated in Peutz-Jeghers syndrome (PJS) and various sporadic cancers. Although LKB1 encodes a kinase that possesses multiple functions, no individual hypothesis posed to date has convincingly explained how loss of LKB1 contributes to carcinogenesis. In this report we demonstrated that LKB1 maintains genomic stability through the regulation of centrosome duplication. We found that LKB1 colocalized with centrosomal proteins and was situated in the mitotic spindle pole. LKB1 deficiency-induced centrosome amplification was independent of AMP-activated protein kinase (AMPK), a well-defined substrate of LKB1. Cells lacking LKB1 exhibited an increase in phosphorylated and total Polo-like kinase 1 (PLK-1), NIMA-related kinase 2 (NEK2), and ninein-like protein (NLP). Overexpression of active PLK1 (T210D) reversed the inhibition of LKB1 on centrosome amplification. In contrast, depletion of PLK1 with siRNA or suppression of PLK1 kinase activity with BTO-1 (5-Cyano-7-nitro-2-benzothiazolecarboxamide-3-oxide) abrogated LKB1 deficiency-induced centrosome amplification. We further characterized that LKB1 phosphorylated and activated AMPK-related kinase 5 (NUAK1 or ARK5) that in turn increased the phosphorylation of MYPT1, enhanced the binding between MYPT1-PP1 and PLK1, and conferred an effective dephosphorylation of PLK1. More importantly, we noted that LKB1-deficient cells exhibited multiple nuclear abnormalities, such as mitotic delay, binuclear, polylobed, grape, large, and micronuclear. Immediate depletion of LKB1 resulted in the accumulation of multiploidy cells. Expression of LKB1 is reversely correlated with the levels of PLK1 in human cancer tissues. Thus, we have uncovered a novel function of LKB1 in the maintenance of genomic stability through the regulation of centrosome mediated by PLK1.

Piperlongumine induces autophagy by targeting p38 signaling.

Piperlongumine (PL), a natural product isolated from the plant species Piper longum L., can selectively induce apoptotic cell death in cancer cells by targeting the stress response to reactive oxygen species (ROS). Here we show that PL induces cell death in the presence of benzyloxycarbonylvalyl-alanyl-aspartic acid (O-methyl)-fluoro-methylketone (zVAD-fmk), a pan-apoptotic inhibitor, and in the presence of necrostatin-1, a necrotic inhibitor. Instead PL-induced cell death can be suppressed by 3-methyladenine, an autophagy inhibitor, and substantially attenuated in cells lacking the autophagy-related 5 (Atg5) gene. We further show that PL enhances autophagy activity without blocking autophagy flux. Application of N-acetyl-cysteine, an antioxidant, markedly reduces PL-induced autophagy and cell death, suggesting an essential role for intracellular ROS in PL-induced autophagy. Furthermore, PL stimulates the activation of p38 protein kinase through ROS-induced stress response and p38 signaling is necessary for the action of PL as SB203580, a p38 inhibitor, or dominant-negative p38 can effectively reduce PL-mediated autophagy. Thus, we have characterized a new mechanism for PL-induced cell death through the ROS-p38 pathway. Our findings support the therapeutic potential of PL by triggering autophagic cell death.