Data_Sheet_1_Pathogenic bacteria significantly increased under oxygen depletion in coastal waters: A continuous observation in the central Bohai Sea.docx

The spread of pathogenic bacteria in coastal waters endangers the health of the local people and jeopardizes the safety of the marine environment. However, their dynamics during seasonal hypoxia in the Bohai Sea (BHS) have not been studied. Here, pathogenic bacteria were detected from the 16S rRNA gene sequencing database and were used to explore their dynamics and driving factors with the progressively deoxygenating in the BHS. Our results showed that pathogenic bacteria were detected in all samples, accounting for 0.13 to 24.65% of the total number of prokaryotic sequences in each sample. Pathogenic Proteobacteria was dominated in all samples, followed by Firmicutes, Actinobacteria, Tenericutes, and Bacteroidetes, etc. β-diversity analysis showed that pathogenic bacteria are highly temporally heterogeneous and regulated by environmental factors. According to RDA analysis, these variations may be influenced by salinity, ammonia, DO, phosphate, silicate, and Chl a. Additionally, pathogenic bacteria in surface water and hypoxia zone were found to be significantly separated in August. The vertical distribution of pathogenic bacterial communities is influenced by several variables, including DO and nutrition. It is noteworthy that the hypoxia zones increase the abundance of certain pathogenic genera, especially Vibrio and Arcobacter, and the stability of the pathogenic bacterial community increased from May to August. These phenomena indicate that the central Bohai Sea is threatened by an increasingly serious pathogenic community from May to August. And the developing hypoxia zone in the future may intensify this phenomenon and pose a more serious threat to human health. This study provides new insight into the changes of pathogenic bacteria in aquatic ecosystems and may help to make effective policies to control the spread of pathogenic bacteria.</p

IMQ Induced K14-VEGF Mouse: A Stable and Long-Term Mouse Model of Psoriasis-Like Inflammation

<div><p>An imiquimod (IMQ) induced wild type (WT) mouse can mimic some features of psoriasis, such as thickened skin, abnormal keratinocyte-related proteins, infiltration of inflammatory cells and pro-inflammatory cytokines. This model is a prevalent model that is widely used in the study of psoriasis. However, skin inflammation decreases during the eighth day when IMQ is given to WT mice, which may result in false results when evaluating the pharmacodynamics effects of a drug. To extend the timeliness and inherit the advantages of this model, we applied IMQ to the skin of 8-week-old homozygous K14-VEGF mice to investigate whether IMQ can prolong mice ear inflammation. In our experiments, we found that, compared to the IMQ induced WT mice model, the IMQ induced K14-VEGF mice have serious skin inflammation, even on the fourteenth day. We also evaluated the stability of skin inflammation at days 8, 10, and 13, and the inflammatory situation remained stable in the skin. This research intends to improve the existing model, and we hypothesize that the IMQ induced K14-VEGF mouse will become a practical mouse model in psoriasis research.</p></div

Curcumin inhibited IMQ-induced psoriasis-like inflammation (N = 6).

<p>A, The thickness of the right ear skin was measured on the days indicated (Mean±SD). B, Erythema, scaling and thickness of the right ear skin was scored on the indicated days with a scale from 0 to 4. The cumulative score is presented (Mean±SD). C, H&E staining of the mouse ear skin of different treatment groups (200×). Arrowheads denote the inside of the ear skin. CUR, curcumin; CLO, clobetasol.</p

Inflammatory changes on days 8–14 in the IMQ-K14 mice.

<p>(a) Changes in ear thickness. n = 6 (b) mRNA level changes of psoriasis-related cytokines, n = 6. (c) Protein level changes of IL-17 and IL-23, n = 3.</p

The possible targets impacted by curcumin in the IL-23/IL-17A axis.

<p><b>CUR, curcumin.</b></p

CLSM images of female <i>S</i>. <i>japonicum</i> from single- and double-sex infections.

<p><b>A.</b> 18SSI with a small and incipient ovary.<b>B.</b>18DSI with a small and incipient ovary that contains a few oogonia, <b>C.</b> 23SSI with an elongated and large ovary that contains a few oogonia. <b>D.</b> 23DSI with a mature and large ovary that contains numerous mature and immature oocytes. OVA, ovary; Scale bars, 25 μm.</p

Immunohistochemical staining for TCR γδ, CCR6 and RORγ in skin.

<p>A, TCR γδ, CCR6 and RORγ immunohistochemical staining was examined in samples of differently treated mouse ear skin (N = 6). B, The mean number of TCR γδ-positive cells per high power field (N = 6). The differences between groups and between any two groups were significant (Kruskal-Wallis test/Wilcoxon Scores test, <i>P</i><0.05). C, CCR6 and RORγ expression was detected using western blotting. CCR6 expression was increased in IMQ-treated mice and was inhibited by both curcumin and clobetasol. The results shown are representative of 3 independent experiments. CUR, curcumin; CLO, clobetasol.</p

IMQ induced a longer skin inflammation period.

<p>(a) mRNA expression of some pro-inflammatory cytokines in the mice skin, n = 6. (b) IHC staining of CCR6⁺ cells in the control group and IMQ-K14 mice on the 14th day. Arrows indicate the location of the CCR6⁺ cells. (c) IHC staining of CD11c⁺ cells in the control group and IMQ-K14 mice on the 14th day. Arrows indicate the location of the CD11c⁺ cells. CH: 8-week-old K14-VEGF mice; IV₁₅: IMQ-K14 14 days.</p

Motif analysis of 24 upregulated ribosomal genes in 23DSI.

<p><b>A.</b> A common motif was found in 11 upregulated ribosomal genes. <b>B.</b> Another motif in other upregulated ribosomal genes. The sequence and site of the two motifs are showed.</p

Differential expression of ribosomal genes in female <i>S</i>. <i>japonicum</i> of single- and double-sex female worms at 18 d and 23 d post-infection.

<p>Pathway analysis of differentially expressed ribosomal genes <b>A.</b> between 23SSI and 18SSI, <b>B.</b> between 18SSI and 18DSI, <b>C.</b> between 23DSI and 18DSI, and <b>D.</b> between 23DSI and 23SSI.Red, upregulated genes; Green, downregulated genes. <b>E.</b> Results of the real-time PCR analysis revealed that the differentially expressed ribosomal genes, such as Sjc_0066600|Sjp_0066600|SJC_S000839.242218,Sjc_0000880|Sjp_0000880|SJC_S000001.1265306,Sjc_0044770|Sjp_0044770|SJC_S000379.21600, showed similar expressions with those found in Solexa analysis. Error bars represent standard deviation obtained from triplicates.(*P<0.05).</p