Comparative characterization of microRNAs of Schistosoma japonicum from SCID mice and BALB/c mice: Clues to the regulation of parasite growth and development.

Schistosomiasis, caused by a parasite with a wide range of mammalian hosts, remains one of the most prevailing parasitic diseases in the world. While numerous studies have reported that the growth and reproduction of schistosomes in immunodeficient mice was significantly retarded, the underlying molecular mechanisms have yet to be revealed. In this study, we comparatively analyzed the microRNA expression of Schistosoma japonicum derived from SCID and BALB/c mice on the 35th day post-infection by high-throughput RNA sequencing as prominent morphological abnormalities had been observed in schistosomes from SCID mice when compared with those from BALB/c mice. The results revealed that more than 72% and 61% of clean reads in the small RNA libraries of female and male schistosomes, respectively, could be mapped to the selected miRs in the miRBase or the sequences of species-specific genomes. Further analysis identified 122 miRNAs using TPM >0.01 as the threshold value, including 75 known and 47 novel miRNAs, 96 of which were commonly expressed across all the four tested schistosome libraries. Comparative analysis of the libraries of schistosomes from SCID and BALB/c mice identified 15 differentially expressed miRNAs (5 up-regulated and 10 down-regulated) among females and 16 among males (9 up-regulated and 7 down-regulated). Integrated analysis of the two sets of differentially expressed miRNAs of female and male worms identified 2 miRNAs (sja-miR-3488 and sja-miR-novel_29) that overlapped between female and male datasets. Prediction of miRNA targets and Gene Ontology (GO) term enrichment analysis of the predicted target genes revealed that these genes were involved in some important biological processes, such as nucleic acid metabolic process, macromolecule modification, and cellular aromatic compound metabolic process. The predicted target genes were further matched to the differentially expressed genes in male and female schistosomes from the above two hosts, obtaining 7 genes that may be responsible for regulating the growth, development and sex maturation of schistosomes. Taken together, this study provides the first identification of differentially expressed miRNAs in schistosomes from SCID and BALB/c mice. These miRNAs and their predicted target mRNAs are probably involved in the regulation of development, growth, and maturation of schistosomes. Therefore, this study expands our understanding of schistosome development regulation and host-parasite relationship, and also provides a valuable set of potential anti-schistosomal targets for prevention and control of schistosomiasis.

Strong Periodic Tendency of Trivalent Lanthanides Coordinated with a Phenanthroline-Based Ligand: Cascade Countercurrent Extraction, Spectroscopy, and Crystallography.

Phenanthroline-diamide ligands have been reported in the selective separation of actinides over Eu(III); on the contrary, relevant basic coordination chemistry studies are still limited, and extraction under actual application conditions is rarely involved. In this work, N,N'-diethyl-N,N'-ditolyl-2,9-diamide-1,10-phenanthroline [Et-Tol-DAPhen (L)] was applied to explore the coordination performance of lanthanides in simulative high-level liquid waste. For the first time, cascade countercurrent extraction was conducted with Et-Tol-DAPhen as the extractant, which reveals the periodic tendency of the extraction efficiency of lanthanides to decrease gradually as the atomic number increases. Comparison of elements with similar radii verifies the hypothesis that the increase in the atomic number leads to a decrease in the ionic radius, thus reducing the coordination and extraction capacity of ligands. Slope analysis, electrospray ionization mass spectrometry, and ultraviolet-visible titration results show that the ligand forms 1:1 and 1:2 complexes with lanthanides and the coordination ability follows the tendency of extraction efficiency, and the first crystal structures of Lns(III) with a phenanthroline-diamide ligand, i.e., [LaL(NO3)3(H2O)] and [LaL2(NO3)2][(NO3)], were obtained, which confirms the conclusions described above. This work promises to enhance our comprehension of the chemical properties of Lns(III) and offer new clues for the design and synthesis of novel separation ligands.

Comparative Transcriptome Analyses of Schistosoma japonicum Derived From SCID Mice and BALB/c Mice: Clues to the Abnormality in Parasite Growth and Development.

Schistosomiasis, caused by the parasitic flatworms called schistosomes, remains one of the most prevailing parasitic diseases in the world. The prodigious oviposition of female worms after maturity is the main driver of pathology due to infection, yet our understanding about the regulation of development and reproduction of schistosomes is limited. Here, we comparatively profiled the transcriptome of Schistosoma japonicum recovered from SCID and BALB/c mice, which were collected 35 days post-infection, when prominent morphological abnormalities could be observed in schistosomes from SCID mice, by performing RNA-seq analysis. Of the 11,183 identified genes, 62 differentially expressed genes (DEGs) with 39 upregulated and 23 downregulated messenger RNAs (mRNAs) were found in male worms from SCID mice (S_M) vs. male worms from BALB/c mice (B_M), and 240 DEGs with 152 upregulated and 88 downregulated mRNAs were found in female worms from SCID mice (S_F) vs. female worms from BALB/c mice (B_F). We also tested nine DEGs with a relatively higher expression abundance in the gonads of the worms (ovary, vitellaria, or testis), suggesting their potential biological significance in the development and reproduction of the parasites. Gene ontology (GO) enrichment analysis revealed that GO terms such as "microtubule-based process," "multicellular organismal development," and "Rho protein signal transduction" were significantly enriched in the DEGs in S_F vs. B_F, whereas GO terms such as "oxidation-reduction process," "response to stress," and "response to DNA damage stimulus" were significantly enriched in the DEGs in S_M vs. B_M. These results revealed that the differential expression of some important genes might contribute to the morphological abnormalities of worms in SCID mice. Furthermore, we selected one DEG, the mitochondrial prohibitin complex protein 1 (Phb1), to perform double-stranded RNA (dsRNA)-mediated RNA interference (RNAi) in vivo targeting the worms in BALB/c mice, and we found that it was essential for the growth and reproductive development of both male and female S. japonicum worms. Taken together, these results provided a wealth of information on the differential gene expression profiles of schistosomes from SCID mice when compared with those from BALB/c mice, which were potentially involved in regulating the growth and development of schistosomes. These findings contributed to an understanding of parasite biology and provided a rich resource for the exploitation of antischistosomal intervention targets.

Comparative Metabonomic Investigations of Schistosoma japonicum From SCID Mice and BALB/c Mice: Clues to Developmental Abnormality of Schistosome in the Immunodeficient Host.

The growth and development of schistosome has been affected in the immunodeficient hosts. But it remains unresolved about the molecular mechanisms involved in the development and reproduction regulation of schistosomes. This study tested and compared the metabolic profiles of the male and female Schistosoma japonicum worms collected from SCID mice and BALB/c mice at 5 weeks post infection using liquid chromatography tandem mass spectrometry (LC-MS/MS) platform, in which the worms from SCID mice were the investigated organisms and the worms from BALB/c mice were used as the controls. There were 1015 ion features in ESI+ mode and 342 ion features in ESI- mode were identified after filtration by false discovery rate. Distinct metabolic profiles were found to clearly differentiate both male and female worms in SCID mice from those in BALB/c mice using multivariate modeling methods including the Principal Component Analysis (PCA), Partial Least Squares Discriminant Analysis (PLS-DA), and Orthogonal Partial Least Squares Discriminant Analysis (OPLS-DA). There were more differential metabolites in female worms than in male worms between SCID mice and BALB/c mice. And common and uniquely perturbed metabolites and pathways were identified among male and female worms from SCID mice when compared with BALB/c mice. The enriched metabolite sets of the differential metabolites in male worms between SCID mice and BALB/c mice included bile acid biosynthesis, taurine and hypotaurine metabolism, sphingolipid metabolism, retinol metabolism, purine metabolism, etc. And the enriched metabolite sets of differential metabolites in female worms included retinol metabolism, alpha linolenic acid and linoleic acid metabolism, purine metabolism, sphingolipid metabolism, glutamate metabolism, etc. Further detection and comparison in transcript abundance of genes of the perturbed retinol metabolism and its associated meiosis process in worms identified clues suggesting accumulated retinyl ester and perturbed meiotic process. These findings suggested an association between the schistosome with retarded growth and development in SCID mice and their perturbed metabolites and metabolic pathways, and provided a new insight into the growth and development regulation of S. japonicum worms from the metabolic level, which indicated great clues for discovery of drugs or vaccines against the parasites and disease with more researches.

[The Effect of Lipopolysaccharide-Induced Mouse B Cell Activation on Schistosoma japonicum Development].

OBJECTIVE: To investigate the effect of lipopolysaccharide (LPS)-induced B cell activation on the development of Schistosoma japonicum. METHODS: Eighteen BALB/c nude mice deficient in T cells and 23 BALB/c SCID mice deficient in T and B cells were used in this study. Each was infected with 30 ± 1 S. japonicum cercariae. The nude (n=9, NL group) and SCID (n=12, SL group) mice then received 2-3 (every two weeks) intraperitoneal injections with LPS (100 µg/mL, 0.2 mL for each mouse). The remaining nude(n=9, N group) and SCID (n=11, S group) mice received PBS injection as control. The mice were sacrificed on days 28 and 36 after infection (n=4/5, 4/5, 5/6, 6/6 for N, NL, S and SL groups, respectively), and adult worms were collected by hepatic portal vein perfusion. The collecting rate of the adult worms was calculated, the body-length measured, and pairs of worms recorded. The liver tissue was collected and digested with 5% KOH, and the number of eggs per gram of liver tissue was calculated. The levels of TGF-ß, IFN-γ and IL-10 in peripheral blood were evaluated. Spleen cell suspension was prepared for detecting the proportion of regulatory B cells (Bregs) in splenic lymphocytes. RESULTS: On day 28 after infection, the body-lengths of male worms in NL and N groups were (7.65±2.85) mm and (5.28±1.64) mm (P<0.01), and those of female worms were (9.64±1.99) mm and (7.49±1.63) mm (P<0.01), respectively. On day 36 after infection, the number of eggs per gram of liver tissue was significantly higher in the NL group than in the N group (1 088±297 vs 715±404, P<0.05), and significantly lower in the SL group than in the S group (217±33 vs 573±160, P<0.01). The proportions of CD(hi)CD5(+)CD19(+) Bregs in N group on days 28 and 36 after infection were (12.73±0.96)% and (37.15±3.04)% (P<0.05), respectively, with no significant difference with that of NL group. The serum levels of TGF-ß and IFN-y on day 28 after infection were significantly different between N and NL groups (TGF-ß, 101.75±46.72 vs 260.90±45.34 pg/mL; IFN-y, 7.91±1.62 vs 14.11±3.72 pg/mL, both P<0.01). Similarly, significant difference was found for the plasma level of IL-10 on day 36 after infection between the S and N groups (41.85±3.14 vs 66.25±4.16 pg/mL, P<0.01), and between the SL and NL groups (44.48±3.87 vs 72.22±17.76 pg/mL, P<0.01), but not between the LPS groups and the control groups. CONCLUSION: LPS can induce the release of cytokines (e.g. TGF-ß) from B cells of mice infected with S. japonium, to facilitate the early development of adult female and male worms.

[Research progress of immunoregulation of B cells to schistosome infection].

Recent studies found that B cell subsets and their factors have double effects on anti- and aiding schistosome infection. This article summarizes the research progress of positive and negative immunoregulation of schistosome infection involving B lymphocytes, antibody and regulatory B cells (Bregs) relating cytokines (IL-10, IL-7 and TGF-ß).

[Blood pressure variation of hypertensive diabetic nephropathy patients undergoing peritoneal dialysis].

OBJECTIVE: To investigate the variance of blood pressure of hypertensive diabetic patients undergoing continuous ambulatory peritoneal dialysis (CAPD). METHODS: Twenty hypertensive CAPD patients older than 40 years with diabetic nephropathy (DN-PD group) and twenty patients with chronic glomerular nephritis (CGN-PD group) were recruited. Peritoneal status and dialysis adequacy of the patients in the two groups were calculated using PD Adequest. All patients were given 24-hour ambulatory blood pressure monitoring (ABPM). Parameters of blood pressure variation were calculated and compared between the two groups, which included 24 h systolic and diastolic blood pressure variability (24 h SBPV/DBPV) and coefficient of variation (24 h SBPCV/24 h DBPCV), daytime systolic anid diastolic blood pressure variability (dSBPV/ DBPV) and coefficient of variation (dSBPCV/dDBPCV), and night time systolic and diastolic blood pressure variability (nSBPV/ DBPV) and coefficient of variation (nSBPCV/nDBPCV). RESULTS: No significant differences in clinical characteristics were found between the two groups of patients except for fast glucose. No significant differences in average systolic and diastolic blood pressures, average piulse pressure and mean 24 h, daytime, and nighttime arterial pressures were found between the two groups. However, the DN-PD group had significantly higher 24 h SBPV, 24 h SBPCV, dSBPV and dSBPCV than the CGN-PD group (P < 0.05). CONCLUSION: Hypertensive diabetic nephropathy patients undergoing peritoneal dialysis have greater blood pressure variance than those with hypertensive chronic glomerular nephritis, despite a similar result of blood pressure control.