USP13 promotes deubiquitination of ZHX2 and tumorigenesis in kidney cancer.

Clear cell renal cell carcinoma (ccRCC) is characterized by the loss of tumor suppressor Von Hippel Lindau (VHL) function. VHL is the component of an E3 ligase complex that promotes the ubiquitination and degradation of hypoxia inducible factor α (HIF-α) (including HIF1α and HIF2α) and Zinc Fingers And Homeoboxes 2 (ZHX2). Our recent research showed that ZHX2 contributed to ccRCC tumorigenesis in a HIF-independent manner. However, it is still unknown whether ZHX2 could be modified through deubiquitination even in the absence of pVHL. Here, we performed a deubiquitinase (DUB) complementary DNA (cDNA) library binding screen and identified USP13 as a DUB that bound ZHX2 and promoted ZHX2 deubiquitination. As a result, USP13 promoted ZHX2 protein stability in an enzymatically dependent manner, and depletion of USP13 led to ZHX2 down-regulation in ccRCC. Functionally, USP13 depletion led to decreased cell proliferation measured by two-dimensional (2D) colony formation and three-dimensional (3D) anchorage-independent growth. Furthermore, USP13 was essential for ccRCC tumor growth in vivo, and the effect was partially mediated by its regulation on ZHX2. Our findings support that USP13 may be a key effector in ccRCC tumorigenesis.

Resilience of fungal flora in bauxite residues amended with organic matter and vermiculite/fly ash.

The fungal community and soil geochemical, physical and biological parameters were analyzed, respectively, in bauxite residues (BRs) treated with organic matter and vermiculite/fly ash by phylogenetic analysis of ITS-18 S rRNA, community level physiological profiles (CLPP) and so on. The results indicated that after amendment of the BR, microbial utilization of carbohydrates and their enzyme activities were significantly increased, but fungal compositions at the phylum level were similar and dominated by the phylum of Ascomycota (82.05-98.96%, RA: relative abundance) after one year of incubation. The fungal taxa in the amended BR treatments, however, show significantly less alpha and beta diversity compared with the reference soils, although they still harbor a substantial novel taxon. The combined amendment of organic matter (OM) and vermiculite/fly ash significantly increases the fungal taxa at the genus and species level compared with solely OM amendment. The results of the following canonical correspondence analysis found that, over 90% variation of the fungal community could be explained by pH, OM and mean weight diameter (MWD) of aggregates; but the biological indicators, including urease (UR), dehydrogenase (DHA) and the value of average well color development (AWCD) could explain only 50% variation of the fungal flora in BRs. This paper indicated that resilience of fungal community in BRs was positively correlated with the BRs' improvement in fertility as well as biogeochemical properties, but alkalinity must be firstly decreased to the target level of BRs' rehabilitation.

Insight of soil amelioration process of bauxite residues amended with organic materials from different sources.

It aimed to investigate and evaluate the soil amelioration process of bauxite residues with the amendments of organic materials from different sources. Wheat straw, poultry manure compost, and biosolids were chosen as the added organic materials. A series of essential soil properties were analyzed to evaluate the effects of organic materials on the soil amelioration of bauxite residue. The results indicated that organic amendments could obviously improve the texture of bauxite residues by increasing large aggregates contents, and elevating its organic matter content and fertility level (such as TN and TP). At the same time, organic additions were effective in reducing bauxite residues' salinity as pH, electrical conductivity and sodium content were obviously decreased in all rehabilitated treatments in comparison with control treatment. These improvements created sufficient conditions for a quick recovery of microbial communities in bauxite residues matrix. The maximum microbial biomass C increased to 0.642 g-C·kg-1, and the activities of urease, catalase, and invertase were massively elevated, especially for those after a year of rehabilitation, although alkali-phosphatase was kept a less level compared with other biological parameters. The further principal analysis and cluster analysis indicated that after 1 year of organic amendment, the improved bauxite residues matrix was very close to the reference soil based on the measured soil microbial properties. All the results suggested that organic amendment is an effective way to stimulate the soil amelioration of bauxite residues, and among the three amended organic materials, wheat straw and biosolid were better in improving the abiotic environmental conditions as well as biotic function recovery in soil amelioration of bauxite residue.

A typical 22q11.2 deletion syndrome and pseudohypoparathyroidism: A CARE compliant case report.

RATIONALE: It is rare to find 22q11.2 deletion syndrome with pseudohypoparathyroidism in children. Furthermore, the phenotypic spectrum of this disorder varies widely. PATIENT CONCERNS: A patient was diagnosed with pseudohypoparathyroidism at age 14 years because of convulsions, hypocalcemia, hyperphosphatemia, normal parathyroid hormone levels, and basal ganglia calcifications. Thereafter, the child presented with symptoms of nephrotic syndrome; subsequently, he was diagnosed with nephrotic syndrome at the local hospital. DIAGNOSIS: At our hospital, multiplex ligation-dependent probe amplification confirmed that the patient had 22q11.2 deletion syndrome. INTERVENTIONS: The patient continued to be treated with calcium supplements. OUTCOMES: Seizure activity and proteinuria ceased. LESSONS: Signs of this syndrome include delayed speech development due to velofacial dysfunction, recurrent croup attacks during early childhood due to latent hypocalcemia, and mild dysmorphic features. The findings of this patient indicated that 22q11.2 deletion syndrome may include a wide spectrum of clinical findings and that this diagnosis needs to be considered for all patients presenting with hypocalcemia, regardless of age.

Identification of small-molecule ion channel modulators in C. elegans channelopathy models.

Ion channels are important therapeutic targets, but the discovery of ion channel drugs remains challenging due to a lack of assays that allow high-throughput screening in the physiological context. Here we report C. elegans phenotype-based methods for screening ion channel drugs. Expression of modified human ether-a-go-go-related gene (hERG) potassium channels in C. elegans results in egg-laying and locomotive defects, which offer indicators for screening small-molecule channel modulators. Screening in worms expressing hERGA561V, which carries a trafficking-defective mutation A561V known to associate with long-QT syndrome, identifies two functional correctors Prostratin and ingenol-3,20-dibenzoate. These compounds activate PKCε signaling and consequently phosphorylate S606 at the pore region of the channel to promote hERGA561V trafficking to the plasma membrane. Importantly, the compounds correct electrophysiological abnormalities in hiPSC-derived cardiomyocytes bearing a heterozygous CRISPR/Cas9-edited hERGA561V. Thus, we have developed an in vivo high-throughput method for screening compounds that have therapeutic potential in treating channelopathies.

Multiple effects of Xihuang pill aqueous extract on the Hs578T triple-negative breast cancer cell line.

The management of triple-negative breast cancer (TNBC) is challenging due to the aggressive behavior, lack of therapeutic options and relatively poor prognosis. Xihuang pill (XHP) is a well-known Traditional Chinese Medicine with anticancer activity. The aim of the present study was to investigate whether the aqueous extract of XHP (AEXHP) has anti-proliferative activity against the Hs578T TNBC cell line, and to elucidate its molecular mechanisms of action. First, an MTT assay was used to evaluate the anti-proliferative activity of AEXHP on the Hs578T cell line; furthermore, the cell cycle distribution, mitochondrial membrane potential and apoptotic rate were determined by flow cytometry, and western blot analysis was used to assess the expression of apoptosis and cell cycle regulatory proteins to investigate the mechanisms of action. The results revealed that the cell viability was significantly inhibited by AEXHP in a dose- and time-dependent manner. Apoptosis and mitochondrial membrane potential loss were detected, and after treatment with 4, 8 and 12 mg/ml AEXHP for 24 h, cleaved caspase-3 was 1.70-, 1.81- and 1.84-fold of that of the control, while procaspase-3, procaspase-8, cleaved caspase-8, B-cell lymphoma 2 (Bcl-2), Bcl-2-associated X protein (Bax) and the Bcl-2/Bax ratio were not significantly affected. Cell cycle analysis revealed that treatment with AEXHP led to S-phase arrest of Hs578T cells. Furthermore, AEXHP treatment resulted in decreased expression of cyclin A and cyclin dependent kinase 2 (CDK2), and increased expression of cyclin E and p21Cip1, as compared to the control group. In conclusion, the viability of Hs578T cells was significantly inhibited by AEXHP in a dose- and time-dependent manner, the likely mechanisms of which being induction of apoptosis, probably via the intrinsic, Bcl-2-independent pathway, and cell cycle arrest in S phase due to decreased expression of cyclin A and CDK2, and increased expression of cyclin E and p21Cip1.

Tanshinone IIA increases the bystander effect of herpes simplex virus thymidine kinase/ganciclovir gene therapy via enhanced gap junctional intercellular communication.

The bystander effect is an intriguing phenomenon by which adjacent cells become sensitized to drug treatment during gene therapy with herpes simplex virus thymidine kinase/ganciclovir (HSV-tk/GCV). This effect is reported to be mediated by gap junctional intercellular communication (GJIC), and therefore, we postulated that upregulation of genes that facilitate GJIC may enhance the HSV-tk/GCV bystander effect. Previous findings have shown Tanshinone IIA (Tan IIA), a chemical substance derived from a Chinese medicine herb, promotes the upregulation of the connexins Cx26 and Cx43 in B16 cells. Because gap junctions are formed by connexins, we hypothesized that Tan IIA might increase GJIC. Our results show that Tan IIA increased GJIC in B16 melanoma cells, leading to more efficient GCV-induced bystander killing in cells stably expressing HSV-tk. Additionally, in vivo experiments demonstrated that tumors in mice with 10% HSV-tk positive B16 cells and 90% wild-type B16 cells became smaller following treatment with the combination of GCV and Tan IIA as compared to GCV or Tan IIA alone. These data demonstrate that Tan IIA can augment the bystander effect of HSV-tk/GCV system through increased gap junction coupling, which adds strength to the promising strategy that develops connexins inducer to potentiate the effects of suicide gene therapy.