Identification of biomarkers and potential therapeutic targets of kidney stone disease using bioinformatics.

PURPOSE: Kidney stone disease (KSD) is a common urological disease, but its pathogenesis remains unclear. In this study, we screened KSD-related hub genes using bioinformatic methods and predicted the related pathways and potential drug targets. METHODS: The GSE75542 and GSE18160 datasets in the Gene Expression Omnibus (GEO) were selected to identify common differentially expressed genes (DEGs). We conducted Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses to identify enriched pathways. Finally, we constructed a hub gene-miRNA network and drug-DEG interaction network. RESULTS: In total, 44 upregulated DEGs and 1 downregulated DEG were selected from the GEO datasets. Signaling pathways, such as leukocyte migration, chemokine activity, NF-κB, TNF, and IL-17, were identified in GO and KEGG. We identified 10 hub genes using Cytohubba. In addition, 21 miRNAs were predicted to regulate 4 or more hub genes, and 10 drugs targeted 2 or more DEGs. LCN2 expression was significantly different between the GEO datasets. Quantitative real-time polymerase chain reaction (qRT-PCR) analyses showed that seven hub gene expressions in HK-2 cells with CaOx treatment were significantly higher than those in the control group. CONCLUSION: The 10 hub genes identified, especially LCN2, may be involved in kidney stone occurrence and development, and may provide new research targets for KSD diagnosis. Furthermore, KSD-related miRNAs may be targeted for the development of novel drugs for KSD treatment.

Overexpression of efflux pump genes is one of the mechanisms causing drug resistance in Mycobacterium tuberculosis.

IMPORTANCE: Gene mutations cannot explain all drug resistance of Mycobacterium tuberculosis, and the overexpression of efflux pump genes is considered another important cause of drug resistance. A total of 46 clinical isolates were included in this study to analyze the overexpression of efflux pump genes in different resistant types of strains. The results showed that overexpression of efflux pump genes did not occur in sensitive strains. There was no significant trend in the overexpression of efflux pump genes before and after one-half of MIC drug induction. By adding the efflux pump inhibitor verapamil, we can observe the decrease of MIC of some drug-resistant strains. At the same time, this study ensured the reliability of calculating the relative expression level of efflux pump genes by screening reference genes and using two reference genes for the normalization of quantitative PCR. Therefore, this study confirms that the overexpression of efflux pump genes plays an important role in the drug resistance of clinical isolates of Mycobacterium tuberculosis.

The survival of murine hepatitis virus (a surrogate of SARS-CoV-2) on conventional packaging materials under cold chain conditions.

Introduction: The cold chain conditions have been suggested to facilitate long-distance transmission of SARS-CoV-2, but it is unclear how viable the virus is on cold chain packaging materials. Methods: This study used the MHV-JHM strain of murine hepatitis virus as a model organism to investigate the viability of SARS-CoV-2 on foam, plastic, cardboard, and wood sheets at different temperatures (-40°C, -20°C, and 4°C). In addition, the ability of peracetic acid and sodium hypochlorite to eliminate the MHV-JHM on plastic and cardboard sheets were also evaluated. Results: The results indicate that MHV-JHM can survive on foam, plastic, or cardboard sheets for up to 28 days at -40°C and -20°C, and up to 14 days on foam and plastic surfaces at 4°C. Although viral nucleic acids were still detectable after storing at 4°C for 28 days, the corresponding virus titer was below the limit of quantification (LOQ). Discussion: The study highlights that a positive nucleic acid test result may not indicate that the virus is still viable, and confirms that peracetic acid and sodium hypochlorite can effectively eliminate MHV-JHM on packaging materials under cold chain conditions.

Administration of a glypican-3 peptide increases the infiltration and cytotoxicity of CD8+ T cells against testicular yolk sac tumor, associated with enhancing the intratumoral cGAS/STING signaling.

BACKGROUND: Glypican-3 (GPC3) is highly expressed in testicular yolk sac tumor (TYST). GPC3 has been evaluated as a cancer vaccine for some types of tumors, but little is known on the effects of GPC3 peptide-based therapy on TYST. Here, we evaluated the antitumor effect of GPC3144-152 on TYST and its potential mechanisms. METHODS: GPC3144-152 -specific CD8+ T cells were induced by vaccine immunization and examined by ELISPOT. The CD8+ T cells were purified for testing their cytotoxicity in vitro against TYST cells by CCK-8 and TUNEL assays and in vivo against tumor growth. The influence of GPC3144-152 loading and/or cGAS silencing on the tumor growth, apoptosis and cGAS/STING signaling was tested by immunohistochemistry, immunofluorescence, flow cytometry, and Western blot. RESULTS: Vaccination with GPC3144-152 induced tumor-specific CD8+ T cells that secreted high levels of IFN-γ and granzyme B, and had potent cytotoxicity against TYST in a dose-dependent manner. Adoptive transfer of CD8+ T cells and treatment with GPC3144-152 significantly inhibited the growth of TYST tumors, but less effective for cGAS-silenced TYST tumors in vivo. Treatment with GPC3144-152 enhanced the infiltration of CD8+ T cells into the tumor environment and their cytotoxicity against TYST tumors in vivo by up-regulating granzyme B and IFN-ß expression, but down-regulating GPC3 expression in the tumors. Co-culture of CD8+ T cells with TYST in the presence of exogenous GPC3144-152 enhanced peptide-specific CD8+ T-cell cytotoxicity in vitro, accompanied by enhancing cGAS, γH2AX, TBK1, and IRF3 phosphorylation in TYST cells, but less effective in cGAS-silenced TYST cells. CONCLUSIONS: These data indicated that GPC3 peptide-specific CD8+ T cells had potent antitumor activity against TYST tumor, particularly for combined treatment with the peptide, which was partially dependent on the intratumoral cGAS/STNG signaling. GPC3 peptide vaccine may be valuable for the combination treatment of TYST.

Experimental Investigation on the Impact of Coal Fines Migration on Pores and Permeability of Cataclastic Coal.

During the production process of coalbed methane, the generation and migration of coal fines can obstruct fractures in coal reservoirs and reduce their permeability. In order to investigate the effects of coal fines migration on the porosity and permeability of coal reservoirs, we conducted core water flooding experiments, low-field nuclear magnetic resonance (NMR), and low-temperature N2 adsorption experiments to study the variations in porosity and permeability of cataclastic coal during coal fines migration and the impact of coal fines migration on porosity and permeability. The experimental results reveal that the initial porosity ratio of cataclastic coal exhibits the characteristics of micropore > macropore > transitional pore > mesopore, with the pore types being predominantly fissured. The porosity of pores larger than 1000 nm and those larger than 10,000 nm exhibit consistent trends before and after water flooding, indicating that the blockage or unblocking of pores with radius larger than 10,000 nm by coal fines can also cause blockage or unblocking of some interconnected macropore. The early stage of flooding is the main period for coal fines migration and production in cataclastic coal, during which the mass concentration of coal fines production is higher and some macropores and fractures become blocked, resulting in a larger decrease in porosity. The higher the initial permeability of cataclastic coal samples with a larger end-face fracture density, the more similar the variations in porosity and permeability of pores larger than 10,000 nm during the flooding experiment, indicating that coal fines mainly block interconnected pores and fractures with radius larger than 10,000 nm through migration, thereby reducing permeability. This study provides a theoretical basis for the efficient production of coalbed methane.

The Relationship between Preoperative Urine Culture and Post-Percutaneous Nephrostolithotomy Systemic Inflammatory Response Syndrome: A Single-Center Retrospective Study.

BACKGROUND: To predict the occurrence of systemic inflammatory response syndrome (SIRS) after percutaneous nephrostrolithotomy(PCNL), preoperative urine culture is a popular method, but the debate about its predictive value is ongoing. In order to better evaluate the value of urine culture before percutaneous nephrolithotomy, we conducted a single-center retrospective study. METHODS: A total of 273 patients who received PCNL in Shanghai Tenth People's Hospital from January 2018 to December 2020 were retrospectively evaluated. Urine culture results, bacterial profiles, and other clinical information were collected. The primary outcome observed was the occurrence of SIRS after PCNL. Univariate and multivariate logistic regression analysis was performed to determine the predictive factors of SIRS after PCNL. A nomogram was constructed using the predictive factors, and the receiver operating characteristic (ROC) curves and calibration plot were drawn. RESULTS: Our results showed that there was a significant correlation between positive preoperative urine cultures and the occurrence of postoperative systemic inflammatory response syndrome. Meanwhile, diabetes, staghorn calculi, and operation time were also risk factors for postoperative systemic inflammatory response syndrome. Our results suggest that among the positive bacteria in urine culture before percutaneous nephrolithotomy, Enterococcus faecalis has become the dominant strain. CONCLUSION: Urine culture is still an important method of preoperative evaluation. A comprehensive evaluation of multiple risk factors should be undertaken and heeded to before percutaneous nephrostrolithotomy. In addition, the impact of changes in bacterial drug resistance is also worthy of attention.

A novel prognostic model based on six methylation-driven genes predicts overall survival for patients with clear cell renal cell carcinoma.

Clear cell renal cell carcinoma (ccRCC) is a lethal urological malignancy. DNA methylation is involved in the regulation of ccRCC occurrence and progression. This study aimed to establish a prognostic model based on DNA methylation to predict the overall survival (OS) of patients with ccRCC. To create this model, we used the transcriptome and DNA methylation data of patients with ccRCC from The Cancer Genome Atlas (TCGA) database. We then used the MethylMix R package to identify methylation-driven genes, and LASSO regression and multivariate Cox regression analyses established the prognostic risk model, from which we derived risk scores. We incorporated these risk scores and clinical parameters to develop a prognostic nomogram to predict 3-, 5-, and 7-year overall survival, and its predictive power was validated using the ArrayExpress cohort. These analyses identified six methylation-driven genes (SAA1, FUT6, SPATA18, SHROOM3, AJAP1, and NPEPL1) that produced risk scores, which were sorted into high- and low-risk patient groups. These two groups differed in nomogram-predicted prognosis, the extent of immune cell infiltration, tumor mutational burden, and expected response to additional therapies. In conclusion, we established a nomogram based on six DNA methylation-driven genes with excellent accuracy for prognostic prediction in ccRCC patients. This nomogram model might provide novel insights into the epigenetic mechanism and individualized treatment of ccRCC.

Risk Factors, Prognostic Factors, and Nomograms for Bone Metastasis in Patients with Newly Diagnosed Clear Cell Renal Cell Carcinoma: A Large Population-Based Study.

Background: This study aimed to investigate risk factors and prognostic factors in patients with clear cell renal cell carcinoma (ccRCC) with bone metastasis (BM) and establish nomograms to provide a quantitative prediction of the risk of BM and survival probability. Methods: The clinicopathological characteristics of patients with ccRCC between January 2010 and December 2015 were obtained from the Surveillance, Epidemiology and End Results (SEER) database. Independent factors for BM in ccRCC patients were identified using univariate and multivariate logistic regression analyses. Prognostic factors for predicting cancer-specific death were evaluated using univariate and multivariate analyses based on a competing risk regression model. We then constructed a diagnostic nomogram and a prognostic nomogram. The two nomograms were evaluated using calibration curves, receiver operating characteristic curves, and decision curve analysis. Results: Our study included 34,659 patients diagnosed with ccRCC in the SEER database, with 1,415 patients who presented with bone metastasis. Risk factors for BM in patients with ccRCC included age, stage T, stage N, brain metastasis, liver metastasis, lung metastasis, tumor size, and laterality. Independent prognostic factors for patients with ccRCC patients with BM were Fuhrman grade, tumor size, T stage, N stage, brain metastases, lung metastasis, and surgery. For the diagnostic nomogram, the area under the curve values in the training and testing cohorts were 0.863 (95% CI, 0.851-0.875) and 0.859 (95% CI, 0.839-0.878), respectively. In the prognostic cohort, the area under the curve values for 1-, 2-, and 3-year cancer-specific survival rates in the training cohort were 0.747, 0.774, and 0.780, respectively, and 0.671, 0.706, and 0.696, respectively, in the testing cohort. Through calibration curves and decision curve analyses, the nomograms displayed excellent performance. Conclusions: Several factors related to the development and prognosis of BM in patients with ccRCC were identified. The nomograms constructed in this study are expected to become effective and precise tools for clinicians to improve cancer management.

Serum and Urinary Neutrophil Gelatinase-Associated Lipocalin Levels as Early Markers of the Renal Function in Patients With Urinary Stone-Induced Hydronephrosis.

Introduction: Urinary stones cause hydronephrosis, which leads to kidney function impairment. The serum creatinine level is frequently used as a marker of kidney function. However, in some patients with hydronephrosis, it does not reflect the kidney function changes in the early stages of kidney stone disease. Neutrophil gelatinase-associated lipocalin (NGAL) is a novel indicator of the kidney function. Previous NGAL-related research has focused on its role in acute kidney injury. This study aimed to determine the usefulness of NGAL as an early marker of the kidney function in patients with urinary stones and hydronephrosis. Methods: Eighty-five patients with urinary stones who were admitted to the Shanghai Tenth People's Hospital (USP group) and 65 healthy volunteers (NC group) were recruited. Blood and urine samples collected from the study participants were evaluated using an enzyme-linked immunosorbent assay to determine the NGAL concentration. Data on the height, weight, age, medical history, and blood and urine findings were collected. Computed tomography data were collected from the USP group. Results: Compared to in the NC group, NGAL levels were significantly elevated in the USP group (P < 0.001). However, no significant differences in the NGAL levels were observed among the USP group members with different degrees of hydronephrosis. Furthermore, no significant between-group differences in the creatinine level or the estimated glomerular filtration rate were observed. The areas under the receiver operating characteristic curves for the serum and urinary NGAL levels with hydronephrosis were 92.03 and 99.54%, respectively. The areas under the receiver operating characteristic curves for the serum and urinary NGAL levels with kidney stones were 85.05 and 91.89%, respectively. Conclusion: NGAL is a sensitive indicator of hydronephrosis secondary to urinary stones.

Early Changes and Predictive Value of Serum Histone H3 Concentration in Urosepsis: A Prospective Observational Study.

INTRODUCTION: To assess early changes in serum histone H3 concentration in patients with urosepsis and its predictive ability for the onset of urosepsis. METHODS: A total of 80 patients who underwent percutaneous nephrolithotripsy were enrolled in the study and divided into control and urosepsis groups based on their postoperative outcomes. Serum histone H3 concentrations were detected using an enzyme-linked immunosorbent assay, blood indexes were tested by automatic blood analyzers, and vital signs data were obtained by monitors and manual measurements. These results were correlated with the incidence of postoperative urosepsis. Repeated measurements and receiver operating characteristic curves were employed to analyze early changes and the predictive value of serum histone H3 concentration in urosepsis. RESULTS: Sixteen of the 80 patients (20%) developed urosepsis after surgery. Our data showed significant intra-group differences in terms of postoperative histone H3 concentrations (P < 0.0001) and variation trends (P < 0.0001). Among analyzed blood markers, serum histone H3 concentrations 3 h postoperation [0.825 (95% confidence interval 0.718-0.931, P < 0.0001; cut-off value 256.74 ng/ml, 93.8% sensitivity, 67.2% specificity)] and 6 h post-operation [0.834 (95% CI 0.721-0.947, P < 0.0001, cut-off value 300.875 ng/ml, 68.8% sensitivity, 87.5% specificity)] displayed a higher area under the corresponding receiver operating characteristic curves, indicating that these markers had a decent predictive value for postoperative urosepsis. CONCLUSION: Our study suggests that serum histone H3 concentration is a novel predictor of postoperative urosepsis in patients undergoing percutaneous nephrolithotripsy. The findings of this study can be validated in a larger cohort. CLINICAL TRIAL REGISTRY NUMBER: ChiCTR1800016679.

The reference intervals for thyroid hormones: A four year investigation in Chinese population.

Introduction: The reference intervals (RIs) are of great importance for physicans to determine whether or not an individual is healthy. However, many clinical laboratories in China still adopted the default RI provided by the manufacturers; and these "uncalibrated" RIs might lead to the misdiagnosis of diseases. In the present study, we enroll reference people with the purpose of determining the RIs of serum triiodothyronine (T3), thyroxine (T4), free triiodothyronine (FT3), free thyroxine (FT4) and thyroid-stimulating hormone (TSH) in Chinese population, and explore the possible roles of age and sex on the levels of biomarkers. Methods: Serum samples from 66,609 individuals who met the inclusion criteria were analyzed using an Roche Cobas E 601 hormone analyzer. The dynamic trends of biomarker were visually assessed by their concentrations over age and sex. Specific partitions were determined by the method of Harris and Boyd. RIs, corresponding to the 2.5th and 97.5th percentiles, as well as the 0.5th, 25th, 50th, 75th and 99.5th percentiles were calculated for each reference partition using a non-parametric rank approach. Results: The serum level of T3, T4, FT4 or TSH showed a right-skewed distribution in both males and females while FT3 presented an approximate normal distribution. Females had a higher mode value of serum T3 or T4, but a lower mode value of serum TSH, FT3 or FT4. All five biomarkers did not need age partitioning according to the approach of harris and boyd, while T3 and FT3 need sex partitioning. Conclusions: The present study not only determined the age- and sex-specific trends of the five thyroid hormones, but provided sex-stratified RIs for T3 and FT3, valuably contributing to the current literature and timely evaluation of thyroid health and disease.

Identification and Validation of a Prognostic Signature for Prostate Cancer Based on Ferroptosis-Related Genes.

Ferroptosis, an iron-dependent form of selective cell death, is involved in the development of many cancers. However, ferroptosis related genes (FRGs) in prostate cancer (PCa) are not been well studied. In this study, we collected the mRNA expression profiles and clinical information of PCa patients from TCGA and MSKCC databases. The univariate, LASSO, and multivariate Cox regression analyses were performed to construct a prognostic signature. Seven FRGs, AKR1C3, ALOXE3, ATP5MC3, CARS1, MT1G, PTGS2, and TFRC, were included to establish a risk model, which was validated in the MSKCC dataset. The results showed that the high-risk group was apparently correlated with copy number alteration load, tumor burden mutation, immune cell infiltration, mRNAsi, immunotherapy, and bicalutamide response. Moreover, we found that TFRC overexpression induced the proliferation and invasion of PCa cell lines in vitro. These results demonstrate that this risk model can accurately predict prognosis, suggesting that FRGs are promising prognostic biomarkers and potential drug targets in PCa patients.

miR-101-3p Serves as a Tumor Suppressor for Renal Cell Carcinoma and Inhibits Its Invasion and Metastasis by Targeting EZH2.

BACKGROUND: The role of miRNAs in renal cell carcinoma (RCC) is not certain. We wanted to study the biological functions and potential mechanisms of miR-101-3p in RCC. METHODS: miR-101-3p was inhibited in A498 and OSRC-2 (two RCC cell lines). We studied its effect on cell invasion and proliferation. Target EZH2 of miR-101-3p was designated by different methods, including luciferase functional analysis and Western blotting. The expression level of the target gene in treated cells was quantitatively analyzed by quantitative real-time polymerase chain reaction. In addition, induction of miR-101-3p to prevent tumor formation of A498 cells in mice was further studied. RESULTS: The overexpression of miR-101-3p significantly inhibited the proliferation, migration, and invasion in two RCC cells. Western blotting and luciferase functional analysis indicated that miR-101-3p regulated the expression of EZH2 in two cell lines. Mice inoculated with A498 and OSRC-2 cells transfected with miR-101-3p mimics showed significantly smaller xenografts and weaker EZH2 expression levels than the control group. CONCLUSIONS: miR-101-3p inhibited RCC cell proliferation, migration, and invasion by targeting EZH2.

ADAM10 and ADAM17 cleave PD-L1 to mediate PD-(L)1 inhibitor resistance.

ADAM10 and ADAM17 expression and soluble PD-L1 (sPD-L1) predict poor prognosis in many malignancies, including in patients treated with PD-(L)1 inhibitors. The mechanism of soluble PD-L1 production and its effects are unknown. Here we uncover a novel mechanism of ADAM10- and ADAM17-mediated resistance to PD-(L)1 inhibitors. ADAM10 and ADAM17 cleave PD-L1 from the surface of malignant cells and extracellular vesicles. This cleavage produces an active sPD-L1 fragment that induces apoptosis in CD8 + T cells and compromises the killing of tumor cells by CD8 + T cells. Reduced tumor site PD-L1 protein-to-mRNA ratios predict poor outcomes and are correlated with elevated ADAM10 and ADAM17 expression in multiple cancers. These results may explain the discordance between PD-L1 immunohistochemistry and PD-(L)1 inhibitor response. Thus, including ADAM10 and ADAM17 tissue staining may improve therapy selection. Furthermore, treatment with an ADAM10/ADAM17 inhibitor may abrogate PD-(L)1 inhibitor resistance and improve clinical responses to PD-(L)1 immunotherapy.

Corrigendum to "Targeting B7-H1 (PD-L1) sensitizes cancer cells to chemotherapy" [Heliyon 4 (12) (December 2018) e01039].

[This corrects the article DOI: 10.1016/j.heliyon.2018.e01039.].

Proteasome activator REGγ promotes inflammation in Leydig cells via IkBε signaling.

The development of testicular inflammation affects the normal male reproductive function. The proteasome activator complex subunit 3 (REGγ) has been suggested to regulate experimental colitis. However, to the best of our knowledge, a potential association between REGγ and testicular inflammation has not been demonstrated. The present study successfully established inflammatory models in C57 mice, primary Leydig cells and the TM3 cell line. It was observed that the absence of REGγ conveyed a significantly protective effect toward testosterone secretion in Leydig cells. REGγ deficiency significantly decreased the expression levels of phosphorylated transcription factor p65 and inflammatory factors in testis tissues, primary Leydig cells and the TM3 cell line. Inflammation also upregulated the expression levels of REGγ. Furthermore, the degradation of the nuclear factor light­chain­enhancer of activated B cells (NF­κB) inhibitor ε (IkBε) signaling pathway regulated REGγ and NF­κB expression. Double knockdown of REGγ and IkBε restored the response in wild­type cells to LPS­induced inflammation. In summary, these results demonstrated that REGγ regulates NF­κB activity by specifically degrading IkBε to regulate inflammation in testicular Leydig cells.

Knockdown of LETM1 inhibits proliferation and metastasis of human renal cell carcinoma cells.

The leucine zipper-EF-hand containing transmembrane protein 1 (LETM1) has been reported to serve an important role in a number of human malignancies and is correlated with poor prognosis. However, little is known about the role of LETM1 in renal cell carcinoma (RCC). In the present study, the expression levels of LETM1 were investigated in RCC cell lines (Caki-1, 786-O, OS-RC-2, A498 and ACHN) and the HK-2 normal human renal tubular epithelial cell line. Short interfering RNA (siRNA) was used to knock down the expression of LETM1 in 786-O and A498 cells. The results indicated that the constitutive expression of LETM1 was notably upregulated in RCC cell lines. Knockdown of LETM1 significantly decreased cell proliferation, migration and invasion. Mechanistically, it was revealed that the knockdown of LETM1 expression sharply downregulated the protein expression of ß-Catenin, Cyclin D1 and c-Myc in 786-O and A498 cells. In conclusion, these results suggest that knockdown of LETM1 exhibits tumor suppressive effects, at least in part by controlling the downstream Wnt/ß-Catenin signaling pathway. Therefore, LETM1 may act as a novel therapeutic target for the treatment of RCC.

CYP3A4 and GCK genetic polymorphisms are the risk factors of tacrolimus-induced new-onset diabetes after transplantation in renal transplant recipients.

PURPOSE: We intend to investigate the association between tacrolimus-induced new-onset diabetes after transplantation (NODAT) and polymorphisms of CYP3A4, CYP3A5, ATP-binding cassette transporter sub-family C member 8 (ABCC8), and glucokinase (GCK) in renal transplant recipients. METHODS: Polymorphisms of CYP3A4 *18B, CYP3A5 *3, ABCC8 T-3C, and GCK G-30A were genotyped in 169 renal transplant recipients. Trough concentrations of tacrolimus were detected by an ELISA kit. The relative materials were collected in all patients and volunteers. The association of NODAT and polymorphisms was analyzed. RESULTS: CYP3A4 *18B and GCK G-30A were related to NODAT (p < 0.05). The lower concentration/dose or fasting serum glucose was in CYP3A4 *1/*1 carriers than that in *18B/*18B carriers in all the renal transplant recipients (p < 0.05), respectively. Genotype of ABCC8 T-3C was associated with fasting serum glucose in both NODAT and non-NODAT patients (p < 0.05). Furthermore, family history of DM (OR = 3.734, p = 0.002), concentration/dose above 70 ([ng/mL]/[mg/kg]) (OR = 2.154, p = 0.034) and GCK G-30A (OR = 2.272, p = 0.026) were independently correlated with the incidence of NODAT in logistic regression. CONCLUSIONS: The polymorphisms of CYP3A4 *18B and GCK G-30A were related to NODAT induced by tacrolimus.

Aldosterone Blocks Rat Stem Leydig Cell Development In Vitro.

Aldosterone (ALDO) is a primary endogenous mineralocorticoid, appearing as the main hormone controlling sodium and water homeostasis. Its emerging role in the development of many organs has gained interest over the past few years. In the testis, Leydig cells contain mineralocorticoid receptors and ALDO stimulates androgen synthesis via the mineralocorticoid receptors in rat adult Leydig cells. Although ALDO pharmacologically promoted the Leydig cell function, its role in Leydig cell development was unclear. In the present study, we investigated effects of ALDO on rat stem Leydig cell (SLC) proliferation and differentiation. Using an in vitro culture system of the seminiferous tubules from Leydig cell-depleted testis and EdU (a modified thymidine analog) incorporation into the SLC for flurorescent labeling to judge its DNA synthesis and measurement of medium testosterone production, steroidogenesis-related gene and protein expression, we found that: (1) ALDO suppressed EdU incorporation into SLCs at 100 nM via mineralocorticoid receptor-mediated mechanism and (2) ALDO reduced Leydig cell number. In conclusion, ALDO pharmacologically blocked rat SLC development.

Targeting B7-H1 (PD-L1) sensitizes cancer cells to chemotherapy.

Development of resistance to chemotherapy is a major obstacle in extending the survival of patients with cancer. Although originally defined as an immune checkpoint molecule, B7-H1 (also named as PD-L1 or CD274) was found to play a role in cancer chemoresistance; however, the underlying mechanism of action of B7-H1 in regulation of chemotherapy sensitivity remains unclear in cancer cells. Here we show that development of chemoresistance depends on an increased activation of ERK in cancer cells overexpressing B7-H1. Conversely, B7-H1 knockout (KO) by CRISPR/Cas9 renders human cancer cells susceptible to chemotherapy in a cell-context dependent manner through a reduced activation of p38 MAPK. B7-H1 was found to associate with the catalytic subunit of DNA-dependent protein kinase (DNA-PKcs) and this association promoted or maintained the activation of ERK or p38 MAPK in cancer cells. Importantly, we found that targeting B7-H1 by anti-B7-H1 monoclonal antibody (H1A) increased the sensitivity of human triple negative breast cancer cells to cisplatin therapy in vivo. Our results suggest that targeting B7-H1 by an antibody capable of disrupting B7-H1 signals may be a new approach to sensitize cancer cells to chemotherapy.