Galectin-9 expression clinically associated with mature dendritic cells infiltration and T cell immune response in colorectal cancer.

BACKGROUND: Galectin-9 is a member of the galectin family and has been reported to have a tumor-promoting or antitumor effect in response to the immune microenvironment. However, the immunomodulatory effect of galectin-9 in colorectal cancer (CRC) remains unclear. The antigen presentation and antitumor immune effects of galectin-9 in CRC were examined in this study. METHODS: The expression of galectin-9, dendritic cell markers (CD208 and CD1a), T-cell markers (CD3 and CD8) and mismatch repair proteins (MLH1, PMS2, MSH2, and MSH6) was assessed using immunohistochemistry in CRC samples. The correlation between galectin-9 and immune cells or immunomodulatory factors was also evaluated via multiple gene expression databases. RESULTS: The level of galectin-9 was decreased in mismatch repair-proficient patients compared with mismatch repair-deficient patients (p = 0.0335). GSEA showed that the regulatory mechanism of galectin-9 in CRC was related to a variety of immune pathways. Galectin-9 expression was strongly correlated with immune cell infiltration and immunomodulators (all p < 0.0001). In the relationship between galectin-9 expression and the infiltration of DCs, there was a negative correlation in CD1a + immature DCs (R = -0.263, p = 0.042). A strong positive correlation was observed in CD208 + mature DCs (R = 0.391, p < 0.01). Patients with high galectin-9 expression also exhibited abundant CD8 + T-cell and CD3 + T-cell infiltration. CONCLUSION: Collectively, our findings provide evidence that galectin-9 may increase the antitumor immune response of patients with CRC. DCs play an important role in galectin-9-mediated antitumor immune responses, which provides further insight into the development of immunotherapy.

Reorganization of the Brain Structural Covariance Network in Ischemic Moyamoya Disease Revealed by Graph Theoretical Analysis.

Objective: Ischemic moyamoya (MMD) disease could alter the cerebral structure, but little is known about the topological organization of the structural covariance network (SCN). This study employed structural magnetic resonance imaging and graph theory to evaluate SCN reorganization in ischemic MMD patients. Method: Forty-nine stroke-free ischemic MMD patients and 49 well-matched healthy controls (HCs) were examined by T1-MPRAGE imaging. Structural images were pre-processed using the Computational Anatomy Toolbox 12 (CAT 12) based on the diffeomorphic anatomical registration through exponentiated lie (DARTEL) algorithm and both the global and regional SCN parameters were calculated and compared using the Graph Analysis Toolbox (GAT). Results: Most of the important metrics of global network organization, including characteristic path length (Lp), clustering coefficient (Cp), assortativity, local efficiency, and transitivity, were significantly reduced in MMD patients compared with HCs. In addition, the regional betweenness centrality (BC) values of the bilateral medial orbitofrontal cortices were significantly lower in MMD patients than in HCs after false discovery rate (FDR) correction for multiple comparisons. The BC was also reduced in the left medial superior frontal gyrus and hippocampus, and increased in the bilateral middle cingulate gyri of patients, but these differences were not significant after FDR correlation. No differences in network resilience were detected by targeted attack analysis or random failure analysis. Conclusions: Both global and regional properties of the SCN are altered in MMD, even in the absence of major stroke or hemorrhagic damage. Patients exhibit a less optimal and more randomized SCN than HCs, and the nodal BC of the bilateral medial orbitofrontal cortices is severely reduced. These changes may account for the cognitive impairments in MMD patients.

Performance and mechanism of the in situ restoration effect on VHCs in the polluted river water based on the orthogonal experiment: photosynthetic fluorescence characteristics and microbial community analysis.

Volatile halogenated hydrocarbons (VHCs) attracted many attentions due to its toxicity and persistence in the environment. In this research, a novel in situ ecological restoration reactor was applied to the degradation of VHCs in polluted river water. The optimized working condition adaptation of the in situ restoration technique was evaluated through orthogonal tests. The experiments showed that when the water depth was 0.4 m, the HRT was 5 days, and the current velocity was 1 m/s, the optimal removal efficiency of VHCs in the reactor was achieved. And the removal rates of CHCl3, CCl4, C2HCl3, and C2Cl4 reached 70.27%, 70.59%, 67.74%, and 81.82%, respectively. The results showed that both HRT and water depth were significantly related to the removal efficiency of reactor. The physiological state of the plants was analyzed by fitting rapid light curve (RLC) model, which showed that the accumulation of VHCs inhibited the photosynthetic performance of plants. Moreover, the microbial community structures of fillers were tested by high-throughput sequencing, and the findings supported that the microbial community made a great response to adapt to the changes in environment of the reactor. The relative abundance of Rhodocyclaceae increased slightly, which hinted that it had good adaptability to VHCs in polluted river water. The research results confirmed that in situ ecological restoration reactor was a potential approach for removal VHCs in polluted river water.

Spatiotemporal evolvement and factors influencing natural and synthetic EDCs and the microbial community at different groundwater depths in the Chaobai watershed: A long-term field study on a river receiving reclaimed water.

In this long-term field study, to restore a dried river ecosystem, reclaimed water was used as a supplementary water source. The main aim of this study was to investigate the accumulation and migration potential of EDCs in groundwater during long-term utilization of reclaimed water and the changes in microbial community during the removal of EDCs. A long-term field study was conducted in order to ascertain the temporal and spatial distribution of four selected endocrine-disrupting chemicals (EDCs) in an underground aquifer in the Chaobai watershed, where reclaimed water is the primary water source. Anew, the microbial community structure at different groundwater depths, along with related environmental factors were also determined. Based on the results obtained from this long-term study, it was found that the EDCs in the surface water of the Chaobai river have entered a depth of 80 m in the groundwater aquifers, within a distance of 360 m from the river. The vertical profiles of the concentrations of bisphenol A (BPA), 4-nonylphenol (NP), estrone (E1), and estriol (E3) decreased significantly from the surface to different groundwater depths with first-order attenuation rates of 0.0416, 0.0343, 0.0498, and 0.0173 m-1. The aquifer depth, water temperature, conductivity, and coexisting anions correlated well with the distribution of EDCs in groundwater.

The ubiquitin ligase Ubr2, a recognition E3 component of the N-end rule pathway, stabilizes Tex19.1 during spermatogenesis.

Ubiquitin E3 ligases target their substrates for ubiquitination, leading to proteasome-mediated degradation or altered biochemical properties. The ubiquitin ligase Ubr2, a recognition E3 component of the N-end rule proteolytic pathway, recognizes proteins with N-terminal destabilizing residues and plays an important role in spermatogenesis. Tex19.1 (also known as Tex19) has been previously identified as a germ cell-specific protein in mouse testis. Here we report that Tex19.1 forms a stable protein complex with Ubr2 in mouse testes. The binding of Tex19.1 to Ubr2 is independent of the second position cysteine of Tex19.1, a putative target for arginylation by the N-end rule pathway R-transferase. The Tex19.1-null mouse mutant phenocopies the Ubr2-deficient mutant in three aspects: heterogeneity of spermatogenic defects, meiotic chromosomal asynapsis, and embryonic lethality preferentially affecting females. In Ubr2-deficient germ cells, Tex19.1 is transcribed, but Tex19.1 protein is absent. Our results suggest that the binding of Ubr2 to Tex19.1 metabolically stabilizes Tex19.1 during spermatogenesis, revealing a new function for Ubr2 outside the conventional N-end rule pathway.

Mouse MOV10L1 associates with Piwi proteins and is an essential component of the Piwi-interacting RNA (piRNA) pathway.

Piwi-interacting RNAs (piRNAs) are essential for silencing of transposable elements in the germline, but their biogenesis is poorly understood. Here we demonstrate that MOV10L1, a germ cell-specific putative RNA helicase, is associated with Piwi proteins. Genetic disruption of the MOV10L1 RNA helicase domain in mice renders both MILI and MIWI2 devoid of piRNAs. Absence of a functional piRNA pathway in Mov10l1 mutant testes causes loss of DNA methylation and subsequent derepression of retrotransposons in germ cells. The Mov10l1 mutant males are sterile owing to complete meiotic arrest. This mouse mutant expresses Piwi proteins but lacks piRNAs, suggesting that MOV10L1 is required for piRNA biogenesis and/or loading to Piwi proteins.

Regulation of male fertility by X-linked genes.

Infertility is a worldwide reproductive health problem, affecting men and women about equally. Mouse genetic studies demonstrate that more than 200 genes specifically or predominantly regulate fertility. However, few genetic causes of infertility in humans have been identified. Here, we focus on the regulation of male fertility by X-linked, germ cell-specific genes. Previous genomic studies reveal that the mammalian X chromosome is enriched for genes expressed in early spermatogenesis. Recent genetic studies in mice show that X-linked, germ cell-specific genes, such as A-kinase anchor protein 4 (Akap4), nuclear RNA export factor 2 (Nxf2), TBP-associated factor 7l (Taf7l), and testis-expressed gene 11 (Tex11), indeed play important roles in the regulation of male fertility. Moreover, we find that the Taf7l Tex11 double-mutant males exhibit much more severe defects in meiosis than either single mutant, suggesting that these 2 X-linked genes regulate male meiosis synergistically. The X-linked, germ cell-specific genes are particularly attractive in the study of male infertility in humans. Because males are hemizygous for X-linked genes, loss-of-function mutations in the single-copy X-linked genes, unlike in autosomal genes, would not be masked by a normal allele. The genetic studies of X-linked, germ cell-specific genes in mice have laid a foundation for mutational analysis of their human orthologues in infertile men.

The pluripotency factor LIN28 marks undifferentiated spermatogonia in mouse.

BACKGROUND: Life-long production of spermatozoa depends on spermatogonial stem cells. Spermatogonial stem cells exist among the most primitive population of germ cells - undifferentiated spermatogonia. Transplantation experiments have demonstrated the functional heterogeneity of undifferentiated spermatogonia. Although the undifferentiated spermatogonia can be topographically divided into As (single), Apr (paired), and Aal (aligned) spermatogonia, subdivision of this primitive cell population using cytological markers would greatly facilitate characterization of their functions. RESULTS: In the present study, we show that LIN28, a pluripotency factor, is specifically expressed in undifferentiated spermatogonia (As, Apr, and Aal) in mouse. Ngn3 also specifically labels undifferentiated spermatogonia. We used Ngn3-GFP knockin mice, in which GFP expression is under the control of all Ngn3 transcription regulatory elements. Remarkably, Ngn3-GFP is only expressed in approximately 40% of LIN28-positive As (single) cells. The percentage of Ngn3-GFP-positive clusters increases dramatically with the chain length of interconnected spermatogonia. CONCLUSION: Our study demonstrates that LIN28 specifically marks undifferentiated spermatogonia in mice. These data, together with previous studies, suggest that the LIN28-expressing undifferentiated spermatogonia exist as two subpopulations: Ngn3-GFP-negative (high stem cell potential) and Ngn3-GFP-positive (high differentiation commitment). Furthermore, Ngn3-GFP-negative cells are found in chains of Ngn3-GFP-positive spermatogonia, suggesting that cells in the Aal spermatogonia could revert to a more primitive state.

Inactivation of Nxf2 causes defects in male meiosis and age-dependent depletion of spermatogonia.

In eukaryotes, mRNA is actively transported from nucleus to cytoplasm by a family of nuclear RNA export factors (NXF). While yeast harbors only one such factor (Mex67p), higher eukaryotes encode multiple NXFs. In mouse, four Nxf genes have been identified: Nxf1, Nxf2, Nxf3, and Nxf7. To date, the function of mouse Nxf genes has not been studied by targeted gene deletion in vivo. Here we report the generation of Nxf2 null mutant mice by homologous recombination in embryonic stem cells. Nxf2-deficient male mice exhibit fertility defects that differ between mouse strains. One third of Nxf2-deficient males on a mixed (C57BL/6x129) genetic background exhibit meiotic arrest and thus are sterile, whereas the remaining males are fertile. Disruption of Nxf2 in inbred (C57BL/6J) males impairs spermatogenesis, resulting in male subfertility, but causes no meiotic arrest. Testis weight and sperm output in C57BL/6J Nxf2(-/Y) mice are sharply reduced. Mutant epididymal sperm exhibit diminished motility. Importantly, proliferation of spermatogonia in Nxf2(-/Y) mice is significantly decreased. As a result, inactivation of Nxf2 causes depletion of germ cells in a substantial fraction of seminiferous tubules in aged mice. These studies demonstrate that Nxf2 plays a dual function in spermatogenesis: regulation of meiosis and maintenance of spermatogonial stem cells.

[Treatment of micro-polluted river water by immobilized microorganism technique].

The effect of immobilized microorganism technique on the micro-polluted river water was studied by four kinds of gaia-biological aerated filter (G-BAF), which were formed by special microorganism (BP35) and four different carriers, including haydite, FPUFS, aquamats flexible carrier and artificial aquatic mat. The removal rates of NH4(+) -N, chlorophyll and turbidity were 83.0%-89.0%, 77.5%-89.0% and 84.4%-95.2%, respectively, and they were all higher than the removal rates of COD, UV254 and TP. The FPUFS contained reactive groups, such as hydroxyl, epoxy and acylamide groups, which made FPUFS load a great amount of enzymes and microorganisms. Therefore, the removal rates of pollutants for FPUFS-G-BAF were higher than those for the other three kinds of G-BAF. Hydraulic retention time (HRT) had little effect on the removal rate of NH4(+) -N, while affected the removal rate of COD significantly. When the concentration of dissolved oxygen (DO) increased from < 2 mg/L to > 4 mg/L, for the four kinds of G-BAF, the removal rates of COD and NH4(+) -N increased 11.9%-18.0% and 12.7%-16.1%, respectively. The result of GC-MS showed that the technique of G-BAF could effectively degrade the macro-molecule refractory organics into small-molecule substance.

Meiotic failure in male mice lacking an X-linked factor.

Meiotic silencing of sex chromosomes may cause their depletion of meiosis-specific genes during evolution. Here, we challenge this hypothesis by reporting the identification of TEX11 as the first X-encoded meiosis-specific factor in mice. TEX11 forms discrete foci on synapsed regions of meiotic chromosomes and appears to be a novel constituent of meiotic nodules involved in recombination. Loss of TEX11 function causes chromosomal asynapsis and reduced crossover formation, leading to elimination of spermatocytes, respectively, at the pachytene and anaphase I stages. Specifically, TEX11-deficient spermatocytes with asynapsed autosomes undergo apoptosis at the pachytene stage, while those with only asynapsed sex chromosomes progress. However, cells that survive the pachytene stage display chromosome nondisjunction at the first meiotic division, resulting in cell death and male infertility. TEX11 interacts with SYCP2, which is an integral component of the synaptonemal complex lateral elements. Thus, TEX11 promotes initiation and/or maintenance of synapsis and formation of crossovers, and may provide a physical link between these two meiotic processes.

Mouse TEX15 is essential for DNA double-strand break repair and chromosomal synapsis during male meiosis.

During meiosis, homologous chromosomes undergo synapsis and recombination. We identify TEX15 as a novel protein that is required for chromosomal synapsis and meiotic recombination. Loss of TEX15 function in mice causes early meiotic arrest in males but not in females. Specifically, TEX15-deficient spermatocytes exhibit a failure in chromosomal synapsis. In mutant spermatocytes, DNA double-strand breaks (DSBs) are formed, but localization of the recombination proteins RAD51 and DMC1 to meiotic chromosomes is severely impaired. Based on these data, we propose that TEX15 regulates the loading of DNA repair proteins onto sites of DSBs and, thus, its absence causes a failure in meiotic recombination.

Abnormal sperm in mice lacking the Taf7l gene.

TFIID is a general transcription factor required for transcription of most protein-coding genes by RNA polymerase II. TAF7L is an X-linked germ cell-specific paralogue of TAF7, which is a generally expressed component of TFIID. Here, we report the generation of Taf7l mutant mice by homologous recombination in embryonic stem cells by using the Cre-loxP strategy. While spermatogenesis was completed in Taf7l(-/Y) mice, the weight of Taf7l(-/Y) testis decreased and the amount of sperm in the epididymides was sharply reduced. Mutant epididymal sperm exhibited abnormal morphology, including folded tails. Sperm motility was significantly reduced, and Taf7l(-/Y) males were fertile with reduced litter size. Microarray profiling revealed that the abundance of six gene transcripts (including Fscn1) in Taf7l(-/Y) testes decreased more than twofold. In particular, FSCN1 is an F-action-bundling protein and thus may be critical for normal sperm morphology and sperm motility. Although deficiency of Taf7l may be compensated in part by Taf7, Taf7l has apparently evolved new specialized functions in the gene-selective transcription in male germ cell differentiation. Our mouse studies suggest that mutations in the human TAF7L gene might be implicated in X-linked oligozoospermia in men.

A novel method to synthesize polyaluminum chloride with a membrane reactor.

Al13 or Alb is usually regarded as the most efficient species of polyaluminum chloride (PAC), the performance flocculant for water treatment. This paper was intended to report a new method to synthesize PAC with high content Alb, by using the membrane reactor. NaOH solutions were managed to permeate slowly through the micropores of ultrafiltration membrane into AICl3 solutions under the suitable transmembrane pressure(TMP). Meanwhile NaOH drops size was limited to nano-scale, resulting in dramatical reduction of the characteristic diffusion time and great increment of contact interface between the strong base and Al ions in solution to favor the formation of Al(OH)4-, the precursor of Al13, so few precipitates and much Alb are produced. When the initial concentration of AlCl3/NaOH is 0.40/2.0 (mol/L), MWCO = 10000, TMP = 0.0085 MPa, T = 305 K and B (molar ratio of OH-/Al3+) = 2.25, the quantity of Alb attains about 80%. The results of 27Al-NMR determination showed that the Al13 content is equal to Alb content. And our PAC product has shown better flocculation effects than the commercial product.