Publications by authors named "Junyu Zhang"

71 Publications

Dynamic content changes of cordycepin and adenosine and transcriptome in Cordyceps kyushuensis Kob at different fermentation stages.

Bioprocess Biosyst Eng 2021 Mar 30. Epub 2021 Mar 30.

State Key Laboratory of Microbial Technology, Shandong University, Qingdao, 266237, China.

20% (w/w) Astragali radix was added to the rice medium to cultivate C. kyushuensis Kob. The fermentation product was collected at mycelium stage, coloring stage, stromata-forming initial stage and fruiting body stage of C. kyushuensis Kob. The dynamic content changes of cordycepin and adenosine were detected at different fermentation stages. In the rice medium with Astragalus radix, both cordycepin and adenosine reached the highest content value on the 30th day of fermentation, 17.31 mg/g and 0.94 mg/g, respectively, which were 8.6 times and 2.0 times of that in rice medium at the same stage. At the same time, transcriptomics technology was used to analyze C. kyushuensis Kob during these four periods.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1007/s00449-021-02561-3DOI Listing
March 2021

[Prenatal diagnosis of spinal muscular atrophy using multiple ligation-dependent probe amplification].

Zhonghua Yi Xue Yi Chuan Xue Za Zhi 2021 Mar;38(3):214-218

Department of Reproductive Genetics, International Peace Maternity and Child Health Care Hospital, School of Medicine, Shanghai Jiaotong University, Shanghai 200030, China.

Objective: To carry out prenatal diagnosis for families with high risk for spinal muscular atrophy (SMA) by using multiplex ligation-dependent probe amplification (MLPA).

Methods: Twenty-one families were enrolled. MLPA was used to detect copy numbers of SMN1 and SMN2 genes. Maternal contamination was excluded by using a short tandem repeat method.

Results: For 23 fetuses from the 21 families, 14 were identified as carriers, 1 as SMA patient, and 8 as normal. By linkage analysis of parental samples, three individuals were determined as silent (2+0) carriers.

Conclusion: MLPA can determine the carrier status of SMA. The identification of three silent (2+0) carriers among the 44 parental samples indicated a risk for such families, for which genetic counseling and reproduction guidance should be provided.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.3760/cma.j.cn511374-20200522-00370DOI Listing
March 2021

Overexpression of isochorismate synthase enhances drought tolerance in barley.

J Plant Physiol 2021 May 9;260:153404. Epub 2021 Mar 9.

College of Life Sciences, Zaozhuang University, Zaozhuang, China; State Key Laboratory of Crop Biology, College of Agronomy, Shandong Agricultural University, Taian, Shandong, China. Electronic address:

Isochorismate synthase (ICS) is a key enzyme for the synthesis of salicylic acid (SA) in plants. SA mediates plant responses to both biotic and abiotic stresses. In previous studies, we found that overexpression of ICS (ICS) or suppression of ICS (ICS) affected the host response to Fusarium graminearum in barley. However, whether the barley ICS gene plays a role in adapting to abiotic stresses remains to be determined. In the present study, expression of the ICS gene was upregulated when treated with 20 % PEG6000, and ICS lines were more drought tolerant than wild type (WT) and ICS. In addition, the abscisic acid (ABA) levels in the ICS lines were higher than those in the WT and ICS lines under drought stress. High ABA levels significantly reduced Gs and E, which may impact water retention under drought stress. Under drought conditions, the activity of antioxidant enzymes was significantly higher in the ICS lines, correlating with a lower levels of reactive oxygen species (ROS) and malondialdehyde (MDA). Enhanced antioxidant competence also contributed to drought tolerance in ICS lines. These findings help elucidate the abiotic stress resistance of the ICS pathway in barley.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1016/j.jplph.2021.153404DOI Listing
May 2021

Ligand-Induced Motion and Self-Assembly Pathways between Nanocubes.

J Phys Chem Lett 2021 Mar 4;12(9):2429-2436. Epub 2021 Mar 4.

Instrumental Analysis Center, Laboratory and Equipment Management Department, Huaqiao University, Xiamen 361021, China.

Nanoparticle motion and self-assembly have been regarded as a promising pathway for forming ordered nanostructures. However, the detailed dynamics processes induced by ligand involvement remained poorly understood. Here, we used liquid-cell electron microscopy technology to image the formation of face-to-face Pt cube ordered structures: pairs, linear chains, and squares. The van der Waals interaction between the two neighboring cubes was quantified in real time. Interestingly, the two different formation processes of the square phase were achieved via a rotational and translational method. It is found that the space between two neighboring cubes was the same as the -TEM results. The density functional theory calculation demonstrated that it was attributed to the DMF ligand interactions of the cubes that promoted their face-to-face attachment.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1021/acs.jpclett.1c00254DOI Listing
March 2021

Astaxanthin inhibits microglia M1 activation against inflammatory injury triggered by lipopolysaccharide through down-regulating miR-31-5p.

Life Sci 2021 Feb 4;267:118943. Epub 2021 Jan 4.

The Marine Biomedical Research Institute, Guangdong Medical University, Zhanjiang 524023, China; Southern Marine Science and Engineering Guangdong Laboratory (Zhanjiang), Zhanjiang 524023, China. Electronic address:

Aims: Astaxanthin is a natural carotenoid, can readily cross the blood-brain barrier and exerts a powerful neuroprotective effect. In this study, experiments were performed to explore the underlying molecular mechanisms of which Astaxanthin inhibiting the microglia M1 activation.

Main Methods: BV2 cells and mice were pre-treated with Astaxanthin and treated by Lipopolysaccharide (LPS). The expressions of M1-related factors (pro-inflammatory cytokines and M1 markers) were measured by RT-qPCR and western blot. The target association between miR-31-5p and Numb was explored via luciferase activity assay. MiR-31-5p mimic was transfected into BV2 cells, then the cells were treated with Astaxanthin in combination with LPS. The expression of M1-related factors and Notch pathway-related molecules were measured via RT-qPCR, western blot and immunofluorescence assay.

Key Findings: Precondition of BV2 cells with Astaxanthin inhibited the expression of M1-related factors triggered by LPS. In addition, Astaxanthin decreased the number of Iba1-positive microglia and downregulated the levels of M1-related factors in hippocampus in LPS-treated mice. Further investigation revealed that Astaxanthin-mediated suppression of M1-related factors levels was reversed by miR-31-5p mimic in BV2 cells stimulated by LPS. Subsequently, we verified that miR-31-5p repressed Numb expression by binding to the 3'-UTR of Numb mRNA. Also, Astaxanthin suppressed the expression of Notch1, Hes1 and Hes5 and improved the expression of Numb in BV2 cells challenged by LPS, but this alteration can be reversed by miR-31-5p mimic.

Significance: Our study demonstrated that down-regulating miR-31-5p by Astaxanthin could be a potential therapeutic approach to suppress neuroinflammation via regulating microglia M1 activation.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1016/j.lfs.2020.118943DOI Listing
February 2021

Removal of Zn(II), Mn(II) and Cu(II) by adsorption onto banana stalk biochar: adsorption process and mechanisms.

Water Sci Technol 2020 Dec;82(12):2962-2974

School of Chemistry and Pharmaceutical Science, Guangxi Normal University, Guilin 541004, China.

Low-cost banana stalk (Musa nana Lour.) biochar was prepared using oxygen-limited pyrolysis (at 500 °C and used), to remove heavy metal ions (including Zn(II), Mn(II) and Cu(II)) from aqueous solution. Adsorption experiments showed that the initial solution pH affected the ability of the biochar to adsorb heavy metal ions in single- and polymetal systems. Compared to Mn(II) and Zn(II), the biochar exhibited highly selective Cu(II) adsorption. The adsorption kinetics of all three metal ions followed the pseudo-second-order kinetic equation. The isotherm data demonstrated the Langmuir model fit for Zn(II), Mn(II) and Cu(II). The results showed that the chemical adsorption of single molecules was the main heavy metal removal mechanism. The maximum adsorption capacities (mg·g) were ranked as Cu(II) (134.88) > Mn(II) (109.10) > Zn(II) (108.10)) by the single-metal adsorption isotherms at 298 K. Moreover, characterization analysis was performed using Fourier transform infrared spectroscopy, the Brunauer-Emmett-Teller method, scanning electron microscopy with energy-dispersive X-ray spectroscopy, X-ray diffraction, and X-ray photoelectron spectroscopy. The results revealed that ion exchange was likely crucial in Mn(II) and Zn(II) removal, while C-O, O-H and C = O possibly were key to Cu(II) removal by complexing or other reactions.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.2166/wst.2020.543DOI Listing
December 2020

Preimplantation Genetic Testing for a Chinese Family With X-Linked Lymphoproliferative Syndrome Type 1.

Front Genet 2020 4;11:550507. Epub 2020 Nov 4.

The International Peace Maternity and Child Health Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, China.

Background: X-linked lymphoproliferative disease (XLP) is a rare primary immunodeficiency disorder. We performed experiments based on two strategies of preimplantation genetic testing (PGT) for a family with XLP caused by a mutation in (c.191G > A).

Methods: First, a single-cell polymerase chain reaction (PCR) protocol was established using single lymphocytes. A nested PCR experiment was performed with direct sequencing after whole genome amplification of single cells to assess the accuracy of the genetic diagnosis. Embryos obtained after intracytoplasmic sperm injection were biopsied on day 3 and detected using the established single-cell PCR protocol. In the second PGT cycle, targeted next generation sequencing (NGS) was performed and the single nucleotide polymorphism (SNP) markers flanking were selected to determine the disease-carrying haplotype phase in each embryo.

Result: In the first PGT cycle, six embryos were biopsied. Discounting an embryo from a single failed PCR experiment, five embryos were identified, including three unaffected and two hemizygous. After PCR, one normal embryo was transferred when it was developing into an early blastocyst. Although the ultrasound images indicated a viable singleton pregnancy, the implantation was on the cesarean scar. Therefore, an artificial abortion was performed. In the haplotyping cycle, six embryos were identified to have inherited a haplotype without pathogenic mutations. After the embryo implantation process failed twice, a successful singleton pregnancy was established, and subsequently, a healthy female child was born.

Conclusion: Targeted NGS with haplotyping analysis circumvents the laborious process of multiplex PCR and is more likely to ensure diagnostic accuracy. However, when a genetic recombination occurs close to the site of mutation, confirmed identification using selected SNP markers can be challenging.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.3389/fgene.2020.550507DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7672036PMC
November 2020

Comprehensive preimplantation genetic testing by massively parallel sequencing.

Hum Reprod 2021 Jan;36(1):236-247

The International Peace Maternity and Child Health Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, China.

Study Question: Can whole genome sequencing (WGS) offer a relatively cost-effective approach for embryonic genome-wide haplotyping and preimplantation genetic testing (PGT) for monogenic disorders (PGT-M), aneuploidy (PGT-A) and structural rearrangements (PGT-SR)?

Summary Answer: Reliable genome-wide haplotyping, PGT-M, PGT-A and PGT-SR could be performed by WGS with 10× depth of parental and 4× depth of embryonic sequencing data.

What Is Known Already: Reduced representation genome sequencing with a genome-wide next-generation sequencing haplarithmisis-based solution has been verified as a generic approach for automated haplotyping and comprehensive PGT. Several low-depth massively parallel sequencing (MPS)-based methods for haplotyping and comprehensive PGT have been developed. However, an additional family member, such as a sibling, or a proband, is required for PGT-M haplotyping using low-depth MPS methods.

Study Design, Size, Duration: In this study, 10 families that had undergone traditional IVF-PGT and 53 embryos, including 13 embryos from two PGT-SR families and 40 embryos from eight PGT-M families, were included to evaluate a WGS-based method. There were 24 blastomeres and 29 blastocysts in total. All embryos were used for PGT-A. Karyomapping validated the WGS results. Clinical outcomes of the 10 families were evaluated.

Participants/materials, Setting, Methods: A blastomere or a few trophectoderm cells from the blastocyst were biopsied, and multiple displacement amplification (MDA) was performed. MDA DNA and bulk DNA of family members were used for library construction. Libraries were sequenced, and data analysis, including haplotype inheritance deduction for PGT-M and PGT-SR and read-count analysis for PGT-A, was performed using an in-house pipeline. Haplotyping with a proband and parent-only haplotyping without additional family members were performed to assess the WGS methodology. Concordance analysis between the WGS results and traditional PGT methods was performed.

Main Results And The Role Of Chance: For the 40 PGT-M and 53 PGT-A embryos, 100% concordance between the WGS and single-nucleotide polymorphism (SNP)-array results was observed, regardless of whether additional family members or a proband was included for PGT-M haplotyping. For the 13 embryos from the two PGT-SR families, the embryonic balanced translocation was detected and 100% concordance between WGS and MicroSeq with PCR-seq was demonstrated.

Limitations, Reasons For Caution: The number of samples in this study was limited. In some cases, the reference embryo for PGT-M or PGT-SR parent-only haplotyping was not available owing to failed direct genotyping.

Wider Implications Of The Findings: WGS-based PGT-A, PGT-M and PGT-SR offered a comprehensive PGT approach for haplotyping without the requirement for additional family members. It provided an improved complementary method to PGT methodologies, such as low-depth MPS- and SNP array-based methods.

Study Funding/competing Interest(s): This research was supported by the research grant from the National Key R&D Program of China (2018YFC0910201 and 2018YFC1004900), the Guangdong province science and technology project of China (2019B020226001), the Shenzhen Birth Defect Screening Project Lab (JZF No. [2016] 750) and the Shenzhen Municipal Government of China (JCYJ20170412152854656). This work was also supported by the National Natural Science Foundation of China (81771638, 81901495 and 81971344), the National Key R&D Program of China (2018YFC1004901 and 2016YFC0905103), the Shanghai Sailing Program (18YF1424800), the Shanghai Municipal Commission of Science and Technology Program (15411964000) and the Shanghai 'Rising Stars of Medical Talent' Youth Development Program Clinical Laboratory Practitioners Program (201972). The authors declare no competing interests.

Trial Registration Number: N/A.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1093/humrep/deaa269DOI Listing
January 2021

A rare cardiac phenotype of dextrocardia observed in a fetus with 1p36 deletion syndrome and a balanced translocation: a prenatal case report.

Mol Cytogenet 2020 Nov 16;13(1):48. Epub 2020 Nov 16.

Prenatal Diagnostic Center, International Peace Maternity and Child Health Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, China.

Background: Chromosome 1p36 deletion syndrome is a contiguous genetic disorder with multiple congenital anomalies and mental retardation. It has been emerging as one of the most common terminal deletion syndromes in humans with the rapid utility of microarray analysis. However, the prenatal findings of 1p36 deletion syndrome are still limited. We report a fetus with 1p36 deletion and cardiac phenotype of dextrocardia, combined with a balanced translocation between chromosome 5 and 6. The phenotype of dextrocardia is rarely reported in prenatal 1p36 deletion cases.

Case Presentation: We present a prenatal 1p36 deletion case with congenital heart diseases and single umbilical artery. Fetal echocardiography showed dextrocardia, ventricular septal defect and pericardial effusion. Fetal karyotype revealed a de novo balanced translocation of 46,XY,t(5;6)(q11.2;q23.3). Chromosomal microarray analysis detected a pathogenic deletion in 1p36.21p36.12, with the size of 6.38 Mb. Further whole genome sequencing revealed that the balanced translocation disrupted the EYA4 and ITGA1 genes.

Conclusions: Although congenital heart diseases are very common clinical manifestations among patients with 1p36 deletion, dextrocardia is a quite rare cardiac phenotype. This is the second case with 1p36 deletion and dextrocardia, and the first prenatally diagnosed 1p36 deletion case with dextrocardia. Our case indicates that genes in 1p36 are associated with not only heart structural anomalies, but also cardiac laterality development. Our results also imply that the EYA4 gene disrupted by the balanced translocation might be related with the cardiac development.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1186/s13039-020-00514-1DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7670715PMC
November 2020

Identification of Key Genes and Pathways Associated with Age-Related Macular Degeneration.

Authors:
Junyu Zhang Yu Zhou

J Ophthalmol 2020 21;2020:2714746. Epub 2020 Aug 21.

Sichuan Provincial Key Laboratory for Human Disease Gene Study, Sichuan Provincial People's Hospital, School of Medicine, University of Electronic Science and Technology of China, Chengdu 611731, China.

Age-related macular degeneration (AMD) is the leading cause of severe, permanent vision loss among the elderly in the developed world. The cellular and molecular pathogenesis of initiation and development of AMD remain poorly delineated. The limited resources of the human AMD RPE/choroid tissues impeded the extensive study of the disease. To better understand the molecular and pathway changes in human AMD RPE/choroid tissues, we searched the literature and found three independent studies using high-throughput technology to analyze gene expression in 54 human AMD RPE/choroid tissues and 46 age-matched healthy controls. We downloaded these data, pooled them together, and reanalyzed the difference between molecular and pathways by the Ingenuity Pathway Analysis (IPA) database. Totally, 353 differentially expressed genes (DEGs) were identified, among which 181 genes were downregulated and 172 genes were upregulated in RPE/choroid of AMD patients. Furthermore, several significantly enriched biological processes, including cancer, organismal injury and abnormalities, and ophthalmic disease, were identified associated with these DEGs. By analysis of canonical pathway, the phototransduction pathway and atherosclerosis signaling were the top two significant canonical pathways altered in RPE/choroid tissues in human AMD. As expected, several ophthalmic disease-related molecules, including RHO, PDE6A, 3',5'-cyclic-GMP phosphodiesterase, and G protein alpha, were in the central nodes of disease network. The bioinformatics technology combined with the existing high-throughput data was applied to evaluate the underlying key genes and pathways in human AMD tissues, which may predict downstream and upstream biological processes and identify potential therapeutic intervention targets in human AMD.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1155/2020/2714746DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7456487PMC
August 2020

Clinical features and expression of type I interferon-inducible genes in systemic lupus erythematosus patients harboring rs1143679 polymorphism in China: a single-center, retrospective study.

Clin Rheumatol 2021 Mar 12;40(3):1093-1101. Epub 2020 Aug 12.

Department of Rheumatology and Immunology, Nanjing First Hospital, Nanjing Medical University, No. 68 Changle Road, Nanjing, 210006, China.

Objective: This case-control study aimed to analyze the clinical features and determine the expression of type I interferon-induced genes in systemic lupus erythematosus (SLE) patients harboring the CD11b rs1143679 single-nucleotide polymorphism (SNP) and elucidate whether it is involved in the relapses of SLE.

Methods: One hundred twenty-five relatively inactive SLE patients with SLEDAI scores < 6, including 102 CD11b rs1143679 G allele patients as controls and 23 rs1143679 A allele carriers as cases, were enrolled from the SLE patient specimen bank in the Department of Rheumatology and Immunology. The sample set was retrospectively analyzed for differences in clinical features, and quantitative PCR and Western blot analyses were performed to evaluate the relative expression of type I interferon (IFN)-inducible genes.

Results: The 24-h urinary protein levels in the case group were significantly elevated, and serum C3 levels were significantly reduced compared with those in the control group (P = 0.019 and P = 0.021, respectively). The relative mRNA levels of IFN-inducible genes IFIT1, IFIT4, and ISG15 in the case group were higher than that in the control group (P = 0.0257, 0.0344, and 0.0311, respectively) and matched with the Western blot results.

Conclusions: The relative expression of type I IFN-inducible genes in inactive SLE patients harboring the CD11b rs1143679 polymorphism was higher than that in other lupus patients. These findings suggest that the rs1143679 SNP can precipitate relapses in inactive SLE patients.

Key Points: • The rs1143679 GA genotype was associated with SLE clinical features. • The rs1143679 GA genotype showed higher interferon-inducible gene expression relative to the GG genotype.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1007/s10067-020-05330-xDOI Listing
March 2021

Clozapine in bipolar disorder: A systematic review and meta-analysis.

J Psychiatr Res 2020 06 27;125:21-27. Epub 2020 Feb 27.

McMaster University, Department of Psychiatry and Behavioural Neurosciences, Hamilton, Canada. Electronic address:

Objectives: To assess the clinical efficacy of clozapine in bipolar disorder and its adverse effect profile.

Methods: A literature search with no year and no language restriction was conducted. The search yielded 3858 articles, with 2453 remaining after duplicate removal; 9 were suitable for the systematic review. From the 9 included studies, 3 (100 patients treated with clozapine and 102 patients treated with other antipsychotics) could be included in a meta-analysis to test clozapine efficacy in the treatment of manic episodes.

Results: Clozapine's efficacy was similar to other antipsychotics (Mean difference (MD): 0.03 [95%CI: 0.86-0.92], p = 0.59) in manic episodes. The systematic review also suggested that clozapine is faster at improving symptoms in manic episodes. In addition, two studies included patients with treatment resistant bipolar disorder (TRBD) and showed that clozapine is superior to other treatments for this specific population. Sedation was the most frequent side effect (49.6%), followed by constipation (31.8%) and tachycardia (23.2%).

Conclusion: Clozapine's efficacy was similar to other antipsychotics in manic episodes and is superior to other antipsychotics among TRBD patients.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1016/j.jpsychires.2020.02.026DOI Listing
June 2020

Clinical Interpretation of Sequence Variants.

Curr Protoc Hum Genet 2020 06;106(1):e98

Department of Pathology, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts.

Clinical interpretation of DNA sequence variants is a critical step in reporting clinical genetic testing results. Application of next-generation sequencing technology in molecular genetic testing has facilitated diagnoses of genetic disorders in clinical practice. However, the large number of DNA sequence variants detected in clinical specimens, many of which have never been seen before, make clinical interpretation challenging. Recommendations by the American College of Medical Genetics and Genomics and the Association for Molecular Pathology (ACMG/AMP) have been widely adopted by clinical laboratories around the world to guide clinical interpretation of sequence variants. The ClinGen Sequence Variant Interpretation Working Group and various disease-specific variant curation expert panels have also developed specifications for the ACMG/AMP recommendations. Despite these efforts to standardize variant interpretation in clinical practice, different laboratories may subjectively use professional judgment to determine which criteria are applicable when classifying a variant. In addition, clinicians and researchers who are not familiar with the variant interpretation process may have difficulty understanding clinical genetic reports and communicating the clinical significance of genetic testing results. Here we provide a step-by-step protocol for clinical interpretation of sequence variants, including practical examples. By following this protocol, clinical laboratory geneticists can interpret the clinical significance of sequence variants according to the ACMG/AMP recommendations and ClinGen framework. Furthermore, this article will help clinicians and researchers to understand variant classification in clinical genetic testing reports and evaluate the quality of the reports. © 2020 by John Wiley & Sons, Inc. Basic Protocol: Interpreting the clinical significance of sequence variants Support Protocol: Reevaluating the clinical significance of sequence variants.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1002/cphg.98DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7431429PMC
June 2020

[Expression and correlation analysis of hypoxia inducible factor 1α and autophagy related molecules in rat nucleus pulposus cells under hypoxia].

Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi 2020 Mar;34(3):318-322

Department of Orthopedics and Traumatology, Yijishan Hospital Affiliated to Wannan Medical College, Wuhu Anhui, 241001, P.R.China.

Objective: To investigate the expression and correlation of hypoxia inducible factor 1α (HIF-1α) and autophagy related molecules (Beclin1 and LC3B) in rat nucleus pulposus cells under hypoxia .

Methods: The nucleus pulposus cells were extracted from the nucleus pulposus of healthy adult Sprague Dawley rats and passaged. The 3rd generation cells were identified by HE staining and collagenase type Ⅱ immunofluorescence staining and randomly divided into 4 groups. The cells in group A were cultured for 8 hours under normal oxygen condition (37℃, 5%CO , 20%O ); the cells in group B were cultured for 8 hours under hypoxia condition (37℃, 5%CO , 1%O ); the cells in group C were transfected with HIF-1α-small interfering RNA and cultured for 8 hours under hypoxia condition; and the cells in group D were cultured with autophagy inhibitor 3-MA for 8 hours under hypoxia condition. Western blot and real-time fluorescence quantitative PCR (qRT-PCR) were used to detect the expressions of HIF-1α and autophagy related molecules (Beclin1 and LC3B) in all groups.

Results: HE staining of the 3rd generation nucleus pulposus cells showed that the cytoplasm was light pink and the nucleus was blue black, and the collagenase type Ⅱ immunofluorescence staining was positive. Western blot and qRT-PCR results showed that the relative expressions of HIF-1α, Beclin1, and LC3B proteins and genes in group B were significantly higher than those in group A ( <0.05); the relative expressions of HIF-1α, Beclin1, and LC3B proteins and genes in group C were significantly lower than those in group B ( <0.05). There was no significant difference in the relative expression of HIF-1α protein and gene between groups B and D ( >0.05); while the relative expressions of Beclin1 and LC3B proteins and genes in group D were significant lower than those in group B ( <0.05).

Conclusion: Hypoxia can induce the expressions of HIF-1α and autophagy related molecules (Beclin1 and LC3B) in rat nucleus pulposus cells, and HIF-1α in rat nucleus pulposus cells under hypoxia is related to the expression of autophagy related molecules, that is, down-regulation of HIF-1α can significantly reduce the expression of autophagy related molecules, while the down-regulation of autophagy levels under hypoxia has no or little effect on the expression of HIF-1α.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.7507/1002-1892.201908088DOI Listing
March 2020

Targeted Sequencing and RNA Assay Reveal a Noncanonical Splicing Variant Causing Alagille Syndrome.

Front Genet 2019 24;10:1363. Epub 2020 Jan 24.

International Peace Maternity and Child Health Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, China.

Alagille syndrome (ALGS), as known as congenital arteriohepatic dysplasia, is a rare autosomal dominant multi-systemic disorder. Mutations in or more rarely have been reported as the cause of ALGS. In this study, a 5-year old girl with typical ALGS feature and her pregnant mother came to our reproductive genetics clinic for counseling. We aimed to clarify the genetic diagnosis and provide prenatal genetic diagnosis for the pregnant. Next generation sequencing (NGS)-based multigene panel was used to identify pathogenic variant of the proband. Then the candidate variant was verified by using Sanger sequencing. RNA assay was performed to clarify splicing effect of the candidate variant. Amniocentesis, karyotyping, and Sanger sequencing were performed for prenatal testing. We found a novel noncanonical splicing variant (c.2917-8C > A) in the proband. Peripheral blood RNA assay suggested that the mutant transcript might escape nonsense-mediated messenger RNA (mRNA) decay (NMD) and encode a C-terminal truncated protein. Information of the variant has resulted in a successful prenatal diagnosis of the fetus. Our results clarified the genetic diagnosis of an ALGS patient and ensured utility of prenatal genetic testing.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.3389/fgene.2019.01363DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6993058PMC
January 2020

Genome sequencing analysis of a family with a child displaying severe abdominal distention and recurrent hypoglycemia.

Mol Genet Genomic Med 2020 03 23;8(3):e1130. Epub 2020 Jan 23.

Department of Medicine-Endocrinology, Baylor College of Medicine, Houston, TX, USA.

Background: Germline mutations in PTEN are associated with the PTEN hamartoma tumor syndrome (PHTS), an umbrella term used to describe a spectrum of autosomal-dominant disorders characterized by variable phenotypic manifestations associated with cell or tissue overgrowth. We report a boy who developed severe progressive abdominal distention due to a dramatic adipose mass from the age of 7 months and developed recurrent hypoinsulinemic hypoglycemia that led to seizures at the age of 4 years.

Methods: Trio-based whole-genome sequencing was performed by using blood DNA from the child and his parents. The possible pathogenic variants were verified by Sanger sequencing. Functional characterization of the identified variant was completed by western blot.

Results: The child inherited a single-nucleotide deletion NM_000314.6:c.849delA (p.Glu284Argfs) in the tumor suppressor gene PTEN from his father. The paternal family members have a history of cancer. It is conceivable that PTEN loss-of-function induced the adipose tumor growth and hypoglycemia, although the proband did not meet the usual diagnosis criteria of Cowden syndrome or Bannayan-Riley-Ruvalcaba syndrome that are characterized by germline mutations of PTEN.

Conclusion: This case underlines the variability of phenotypes associated with PTEN germline mutations and provides useful information for diagnosis and genetic counseling of PTEN-related diseases for pediatric patients.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1002/mgg3.1130DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7057095PMC
March 2020

Transcriptome sequencing of adenomyosis eutopic endometrium: A new insight into its pathophysiology.

J Cell Mol Med 2019 12 1;23(12):8381-8391. Epub 2019 Oct 1.

International Peace Maternity and Child Health Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, China.

The eutopic endometrium has been suggested to play a crucial role in the pathogenesis of adenomyosis. However, the specific genes in eutopic endometrium responsible for the pathogenesis of adenomyosis still remain to be elucidated. We aim to identify differentially expressed genes (DEGs) and molecular pathways/networks in eutopic endometrium from adenomyosis patients and provide a new insight into disease mechanisms at transcriptome level. RNA sequencing (RNA-Seq) was performed with 12 eutopic endometrium from adenomyosis and control groups. Differentially expressed genes in adenomyosis were validated by quantitative real-time PCR (qPCR) and immunochemistry. Functional annotations of the DEGs were analysed with Ingenuity Pathway Analysis (IPA). Quantitative DNA methylation analysis of CEBPB was performed with MassArray system. A total of 373 differentially expressed genes were identified in the adenomyosis eutopic endometrium compared to matched controls. Bioinformatic analysis predicted that IL-6 signalling and ERK/MAPK signalling were activated in adenomyosis endometrium. We also found that the increased expression and DNA hypomethylation of CEBPB were associated with adenomyosis. Our results revealed key pathways and networks in eutopic endometrium of adenomyosis. The study is the first to propose the association between C/EBPβ and adenomyosis and can improve the understanding of the pathogenesis of adenomyosis.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1111/jcmm.14718DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6850960PMC
December 2019

The Protein Phosphatase 1 Complex Is a Direct Target of AKT that Links Insulin Signaling to Hepatic Glycogen Deposition.

Cell Rep 2019 09;28(13):3406-3422.e7

State Key Laboratory of Membrane Biology and Tsinghua-Peking Center for Life Sciences, School of Life Sciences, Tsinghua University, Beijing 100084, China. Electronic address:

Insulin-stimulated hepatic glycogen synthesis is central to glucose homeostasis. Here, we show that PPP1R3G, a regulatory subunit of protein phosphatase 1 (PP1), is directly phosphorylated by AKT. PPP1R3G phosphorylation fluctuates with fasting-refeeding cycle and is required for insulin-stimulated dephosphorylation, i.e., activation of glycogen synthase (GS) in hepatocytes. In this study, we demonstrate that knockdown of PPP1R3G significantly inhibits insulin response. The introduction of wild-type PPP1R3G, and not phosphorylation-defective mutants, increases hepatic glycogen deposition, blood glucose clearance, and insulin sensitivity in vivo. Mechanistically, phosphorylated PPP1R3G displays increased binding for, and promotes dephosphorylation of, phospho-GS. Furthermore, PPP1R3B, another regulatory subunit of PP1, binds to the dephosphorylated GS, thereby relaying insulin stimulation to hepatic glycogen deposition. Importantly, this PP1-mediated signaling cascade is independent of GSK3. Therefore, we reveal a regulatory axis consisting of insulin/AKT/PPP1R3G/PPP1R3B that operates in parallel to the GSK3-dependent pathway, controlling glycogen synthesis and glucose homeostasis in insulin signaling.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1016/j.celrep.2019.08.066DOI Listing
September 2019

Enhanced Stabilization and Effective Utilization of Atomic Hydrogen on Pd-In Nanoparticles in a Flow-through Electrode.

Environ Sci Technol 2019 Oct 17;53(19):11383-11390. Epub 2019 Sep 17.

Key Laboratory of Drinking Water Science and Technology, Research Center for Eco-Environmental Sciences , Chinese Academy of Sciences , Beijing 100085 , China.

Surface-adsorbed active species are intermediates with strong activities in heterogeneous catalytic reactions. Effective stabilization of these intermediates is crucial to improve the catalytic performance. Here, we demonstrated highly active bimetallic palladium-indium (Pd-In) nanoparticles (NPs) that can stabilize atomic H* on the surface and show efficient electrocatalytic reduction performance toward bromate. The optimal atomic ratio of Pd to In was investigated with the aim of efficient formation and strong stabilization of H*, thus facilitating the reduction and decontamination of carcinogenic bromate. PdIn was the most active catalyst, with a high rate constant of 0.029 min, whereas the rate constant for monometallic Pd NPs was only 0.009 min. Density functional theory calculations suggest that PdIn NPs decrease the work function and provide strong H* stabilization ability. By employing a flow-through electrode coated with PdIn NPs to enhance the mass transport, the utilization of H* could be boosted and the reduction kinetics increased up to 7.5 times.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1021/acs.est.9b03111DOI Listing
October 2019

Poliovirus receptor CD155 is up-regulated in muscle-invasive bladder cancer and predicts poor prognosis.

Urol Oncol 2020 02 2;38(2):41.e11-41.e18. Epub 2019 Aug 2.

Department of Urology, Fudan University Shanghai Cancer Center, Shanghai, China. Electronic address:

Objective: To investigate the expression pattern of CD155 and evaluate the prognostic value of CD155 in muscle-invasive bladder cancer (MIBC).

Patients And Methods: Immunohistochemical staining of CD155 and survival analysis were conducted on 228 nonmetastatic MIBC patients underwent radical cystectomy in cohorts from Fudan University Shanghai Cancer Center and Zhongshan Hospital. Association of CD155 gene expression with tumor stage and survival were analyzed in TCGA and GSE13507 dataset.

Results: CD155 was significantly up-regulated in MIBC compared to matched normal urothelium and majorly stained on the membrane of tumor cells. In Fudan MIBC cohort, CD155 high expression was significantly correlated with shorter recurrence-free survival (HR = 2.13, P < 0.001) and overall survival (HR = 2.49, P < 0.001). CD155 expression, T stage, and lymph node status were independent factors for predicting survival in multivariate analysis. In TCGA dataset, CD155 high expression was independently associated with shorter overall survival (HR = 1.74, P = 0.001) beyond age, T stage, and lymph node status. Further, explorative analysis in Fudan MIBC cohort showed that adjuvant chemotherapy was associated with longer recurrence-free survival and overall survival in stage III and IV disease with CD155-high tumors.

Conclusions: These findings suggest that CD155 is a robust prognostic factor and may help predict the benefit of adjuvant chemotherapy in MIBC.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1016/j.urolonc.2019.07.006DOI Listing
February 2020

Epigenome-wide association data implicate fetal/maternal adaptations contributing to clinical outcomes in preeclampsia.

Epigenomics 2019 07 16;11(9):1003-1019. Epub 2019 May 16.

International Peace Maternity & Child Health Hospital of China Affiliated to Shanghai Jiao Tong University, Shanghai, 200030, PR China.

To investigate the changes of placental DNA methylome in preeclampsia (PE). We performed an epigenome-wide association study in a Chinese cohort and six published datasets consisting of 335 samples in total. Numerous consistently hypomethylated probes were associated with early-onset PE in different populations, with 2125 reaching epigenome-wide significance. The validated probes were enriched for cytosine-phosphate-guanine dinucleotide (CpG) sites partially methylated and located in genes related to trophoblast fusion. The methylation levels of the validated probes were associated with clinical severity, while the intermediate samples showed antagonistic fetal/maternal outcomes. The DNA methylation patterns of PE and clinically relevant obstetrical syndromes suggested partially common pathophysiologies and complex maternal/fetal adaptive responses contributing to variable clinical outcomes.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.2217/epi-2019-0065DOI Listing
July 2019

Deactivation of Cu-SAPO-34 by urea-related deposits at low temperatures and the regeneration.

J Environ Sci (China) 2019 Jul 18;81:43-51. Epub 2019 Feb 18.

The Key Laboratory of Advanced Materials of Ministry of Education, School of Materials Science and Engineering, Tsinghua University, Beijing 100084, China. E-mail: Graduate School at Shenzhen, Tsinghua University, Shenzhen City 518055, China. Electronic address:

Selective catalytic reduction (SCR) with urea catalyzed by Cu-SAPO-34 is an effective method to eliminate NO from diesel exhaust. However, urea-related deposits may form during cold-start and urban driving due to low exhaust temperatures. The activity of Cu-SAPO-34 at 175°C is significantly degraded by urea exposure, and 300°C is required for regeneration. Through in-situ diffuse reflectance infrared Fourier transform spectroscopy (DRIFTS) and temperature-programmed hydrolysis studies, the dominant stable deposit at 175°C is identified as biuret, which can be eliminated at 300°C. The urea-derived deactivation and regeneration mechanisms of Cu-SAPO-34 were compared with those of anatase-supported catalysts.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1016/j.jes.2019.02.008DOI Listing
July 2019

Improving the Photostability of Ultrasmall Au Clusters via a Combined Strategy of Surface Engineering and Interfacial Modification.

Langmuir 2019 Apr 17;35(17):5728-5736. Epub 2019 Apr 17.

State Key Laboratory of Photocatalysis on Energy and Environment, College of Chemistry , Fuzhou University , Fuzhou 350116 , P. R. China.

Photostability is a critical issue for evaluating the use of photocatalysts to realize large-scale implementation of solar energy conversion. Recently emerged ultrasmall gold (Au) clusters with distinctive physicochemical properties have been regarded as visible-light photosensitizers for photoredox catalysis, whereas the poor stability under visible-light exposure greatly restricts their photocatalytic applications. Herein, we provide a proof-of-concept study on enhancing the photostability of ultrasmall Au clusters via a combined strategy of surface engineering and interfacial modification. The photostability of Au clusters on the surface of TiO nanosheets with less hydroxyl group can be improved to some extent as compared to that on TiO nanoparticles with abundant hydroxyl groups under continuous visible-light irradiation (λ > 420 nm). Moreover, the subsequent modification of branched polyethylenimine (BPEI) between TiO nanosheets and Au clusters further improves their photostability upon light illumination. Consequently, the as-constructed TiO nanosheet-BPEI-Au cluster composites exhibit stable visible-light activity toward Cr(VI) photoreduction. It is hoped that the joint strategy via surface engineering and interfacial modification provides a facile guideline for stabilizing ultrasmall Au clusters toward targeting applications in the photoredox catalysis process.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1021/acs.langmuir.9b00404DOI Listing
April 2019

The effects of perinatal bisphenol A exposure on thyroid hormone homeostasis and glucose metabolism in the prefrontal cortex and hippocampus of rats.

Brain Behav 2019 03 13;9(3):e01225. Epub 2019 Feb 13.

Medical Institute of Developmental Disorders Research, Showa University, Tokyo, Japan.

Introduction: Bisphenol A (BPA) is an endocrine disruptor widely used to manufacture consumer goods. Although the thyroid hormone (TH) disrupting potential of BPA has been thought to be responsible for the neuropsychiatric deficits in the animals that experienced perinatal BPA exposure, the TH availability change at the level of specific brain structures has not been subject to systematic investigation.

Methods: In the present study the impacts of perinatal BPA exposure (0.1 mg/L in drinking water) spanning gestation and lactation on TH homeostasis in the prefrontal cortex (PFC) and hippocampus were assessed in male Sprague-Dawley rats at postnatal day 21 (PND21) and PND90. As TH regulates brain glucose metabolism at multiple levels,the effects of BPA treatment on glucose metabolism in the brain tissues were also assessed in adult rats.

Results: The results showed heterogeneous changes in TH concentration induced by BPA between serum and brain tissues, additionally, in the BPA-treated pups, up-regulated expression of the TH transporter monocarboxylate 8 mRNA at PND21 and increased type 3 iodothyronine deiodinase mRNA expressions at PND21 and PND90 were observed. Meanwhile, decreased glucose metabolism was seen in the PFC and hippocampus, while deficits in locomotor activity, spatial memory and social behaviors occurred in BPA-treated groups.

Conclusion: These data support the concept that the developing brain possesses potent mechanisms to compensate for a small reduction in serum TH, such as serum hypothyrodism induced by BPA exposure, however, the long-term negative effect of BPA treatment on TH homeostasis and glucose metabolism may be attributable to neuropsychiatric deficits after mature.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1002/brb3.1225DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6422808PMC
March 2019

Incorporating non-biological factors into the TNM staging system for better prognostication and decision-making in testicular cancer.

World J Urol 2019 Oct 15;37(10):2165-2173. Epub 2018 Dec 15.

Department of Urology, Fudan University Shanghai Cancer Center, Shanghai, 200032, China.

Background: We combined county-level socioeconomic status (SES), marital status and insurance status to introduce NBF-stage, which were further incorporated into the American Joint Committee on Cancer (AJCC) TNM staging system to generate an integrated staging system for better prognostication and decision-making for testicular cancer patients.

Methods: 15,324 eligible patients diagnosed with primary testicular cancer between January 1, 2007 and December 31, 2015 were strictly selected from the Surveillance, Epidemiology, and End Results (SEER) database. Independent survival predictors were determined based on Cox proportional hazards model. The Kaplan-Meier survival curves were conducted to describe the difference in predicting survival probability and the Multivariate Cox proportion hazard regression analyses were established to compare the cancer-specific survival (CSS) and overall survival (OS) difference among NBF stages or NBF-TNM subgroups.

Results: County-level SES, marital status and insurance status were independent prognostic non-biological factors (NBFs) in our study (P < 0.05). NBF-stage (combination of SES, marital status, and insurance status) was also an independent survival predictor in TC (P < 0.05). NBF1 patients had 167% increased risk of cancer-specific mortality (CSM) as compared to NBF0 patients in testicular cancer (P < 0.01). And NBF0 patients all had a better CSS as compared to NBF1 patients of the same TNM stage both in seminoma and non-seminomatous germ cell tumor (P < 0.05).

Conclusions: Incorporation of NBFs into AJCC TNM staging system in testicular cancer would potentially impact treatment decisions where treatments would not be rendered for a typically curable cancer with multi-modal therapy.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1007/s00345-018-2603-1DOI Listing
October 2019

Evaluation of Proton Therapy Accuracy Using a PMMA Phantom and PET Prediction Module.

Front Oncol 2018 13;8:523. Epub 2018 Nov 13.

Department of Nuclear Medicine, Shanghai Proton and Heavy Ion Center, Fudan University Cancer Hospital, Shanghai, China.

Positron emission tomography (PET) scanning is a widely used method of proton therapy verification. In this study, a proton radiotherapy accuracy verification process was developed by comparing predicted and measured PET data to verify the correctness of PET prediction and was tested at the Shanghai Proton and Heavy Ion Center. Irradiation was performed on a polymethyl methacrylate (PMMA) phantom. There were two dose groups, to which 2 and 4 Gy doses were delivered, and each dose group had different designed dose depths ranging from 5 to 20 cm. The predicted PET results were obtained using a PET prediction calculation module. The measured data were collected with a PET/computed tomography device. The predicted and measured PET data were normalized to similar PET amplitude values before comparison and were compared using depth and lateral profiles for the position error. The error was evaluated at the position corresponding to 50% of the maximum on the PET curves. The mean and standard deviation were calculated based on the data sampled in the scoring area. Gamma index analysis is also applied in the comparison. In the depth comparison, the 2 and 4 Gy dose cases yielded similar mean depth errors between 1 and -1 mm, and the deviation was <2 mm. In the lateral comparison, the 2 Gy cases had a mean lateral error around 1 mm, and the 4 Gy cases had a mean lateral error <1 mm, with a standard deviation <1 mm for both the 2 and 4 Gy cases. All the cases have a gamma passing rate over 95%. The comparison of these PMMA phantom cases revealed good agreement between the predicted and measured PET data, with depth and lateral position errors <2 mm in total, considering the uncertainty. The comparison results demonstrate that the PET predictions obtained in PMMA phantom tests for single proton beam therapy verification are reliable and that the research can be extended to verification in human body treatment with further investigation.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.3389/fonc.2018.00523DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6243057PMC
November 2018

Tumor stroma-infiltrating mast cells predict prognosis and adjuvant chemotherapeutic benefits in patients with muscle invasive bladder cancer.

Oncoimmunology 2018;7(9):e1474317. Epub 2018 Aug 1.

Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Fudan University, Shanghai, China.

Which subgroups patients with muscle-invasive bladder cancer (MIBC) could benefit most from adjuvant chemotherapy (ACT) is blurred. Here we tried to stratify MIBC patients with tumor infiltrating mast cells (TIMs), explore the prognostic and predictive value of TIMs, and provide possible cellular explanations. We selected 259 MIBC patients who underwent radical cystectomy from two independent clinical centers between 2002 and 2014. TIMs were evaluated and prognostic and predictive value was assessed. The CIBERSORT method, Gene Set Enrichment Analysis (GSEA) and differential gene expression analyses were performed to explore the possible cellular mechanisms. TIMs infiltration was distinct between stromal and epithelial area of MIBC specimens. Patients with higher stromal TIMs had a significant worse overall survival and recurrence free survival (HR = 2.228, 95%CI: 1.467-3.550; = 0.001 and HR = 1.984, 95%CI: 1.105-3.374; = 0.016). More importantly, pT2 patients with low stromal TIMs tended to have a lower risk of death and recurrence after ACT (HR = 0.233, 95%CI: 0.020-0.814; = 0.033 and HR = 0.180, 95%CI: 0.022-0.722; = 0.031). A negative correlativity between TIMs and CD8 + T cells was identified on TCGA-BLCA cohort. Immunohistochemistry results validated that high stromal TIMs were negatively correlated with CD8 + T cells (Spearman's rho = -0.215, < 0.001). Differential gene expression suggested that low TIMs might represent a state of immune activation in MIBC. To conclude, high stromal TIMs infiltration was an independent unfavorable prognosticator for MIBC patients. Patients with low stromal TIMs might benefit the most from ACT, especially in pT2 stage.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1080/2162402X.2018.1474317DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6209422PMC
August 2018

Tumor-associated Macrophage-derived Interleukin-23 Interlinks Kidney Cancer Glutamine Addiction with Immune Evasion.

Eur Urol 2019 05 4;75(5):752-763. Epub 2018 Oct 4.

Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Fudan University, Shanghai, China. Electronic address:

Background: Glutamine addiction is a hallmark of clear cell renal cell carcinoma (ccRCC); yet whether glutamine metabolism impacts local immune surveillance is unclear. This knowledge may yield novel immunotherapeutic opportunities.

Objective: To seek a potential therapeutic target in glutamine-addicted ccRCC.

Design, Setting, And Participants: Tumors from ccRCC patients from a Shanghai cohort and ccRCC tumor data from The Cancer Genome Atlas (TCGA) cohort were analyzed. In vivo and in vitro studies were conducted with fresh human ccRCC tumors and murine tumor cells.

Outcome Measurements And Statistical Analysis: Immune cell numbers and functions were analyzed by flow cytometry. Glutamine and cytokine concentrations were determined. Survival was compared between different subpopulations of patients using Kaplan-Meier and Cox regression analyses.

Results And Limitations: We found that in ccRCC, high interleukin (IL)-23 expression was significantly associated with poor survival in both TCGA (overall survival [OS] hazard ratio [HR]=2.04, cancer-specific survival [CSS] HR=2.95; all p<0.001) and Shanghai (OS HR=2.07, CSS HR=3.92; all p<0.001) cohorts. IL-23 blockade prolongs the survival of tumor-bearing mice, promotes T-cell cytotoxicity in in vitro cultures of human ccRCC tumors, and augments the therapeutic benefits of anti-PD-1 antibodies. Mechanistically, glutamine consumption by ccRCC tumor cells results in the local deprivation of extracellular glutamine, which induces IL-23 secretion by tumor-infiltrating macrophages via the activation of hypoxia-inducible factor 1α (HIF1α). IL-23 activates regulatory T-cell proliferation and promotes IL-10 and transforming growth factor β expression, thereby suppressing tumor cell killing by cytotoxic lymphocytes. The positive correlations between glutamine metabolism, IL-23 levels, and Treg responses are confirmed in both TCGA cohort and tumors from Shanghai ccRCC patients. Study limitations include the unclear impacts of glutamine deprivation and IL-23 on other immune cells.

Conclusions: Macrophage-secreted IL-23 enhanced Treg functions in glutamine-addicted tumors; thus, IL-23 is a promising target for immunotherapy in ccRCC.

Patient Summary: In this study, we analyzed the immune components in glutamine-addicted clear cell renal cell carcinoma (ccRCC) tumors from two patient cohorts and conducted both in vitro and in vivo studies. We found that ccRCC tumor cell-intrinsic glutamine metabolism orchestrates immune evasion via interleukin (IL)-23, and IL-23-high patients had significantly poorer survival than IL-23-low patients. IL-23 should thus be considered a therapeutic target in ccRCC, either alone or in combination with immune checkpoint inhibitors.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1016/j.eururo.2018.09.030DOI Listing
May 2019

CD19 tumor-infiltrating B-cells prime CD4 T-cell immunity and predict platinum-based chemotherapy efficacy in muscle-invasive bladder cancer.

Cancer Immunol Immunother 2019 Jan 26;68(1):45-56. Epub 2018 Sep 26.

Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Fudan University, Building 7, No. 130, Dongan Road, Shanghai, 200032, China.

Purpose: CD19 tumor-infiltrating B-cells (CD19 TIB) play a crucial role in tumorigenesis, but their clinical relevance in muscle-invasive bladder cancer (MIBC) remains unknown. This study aimed to investigate the prognostic value of CD19 TIB for post-surgery survival and adjuvant chemotherapy response in MIBC.

Experimental Design: We assessed TIB by immunohistochemical staining of CD19 in 246 MIBC patients from Zhongshan Hospital and Shanghai Cancer Center. We evaluated the survival benefit of platinum-based chemotherapy according to CD19 TIB. The mechanism underlying CD19 TIB antitumor immunity was explored through the Cancer Genome Atlas (TCGA) dataset analysis and an in vitro Ag presentation assay.

Results: CD19 TIB extensively infiltrated into the tumor stroma of MIBC. Adjuvant chemotherapy (ACT) led to a significantly increased benefit in the high CD19 TIB MIBC patients (P = 0.003). In multivariate analysis, high CD19 TIB MIBC patients had significantly longer OS with ACT in the discovery set (HR = 0.487, P = 0.038). TCGA gene expression analyses showed enrichment of adaptive immunity, T-cell-mediated immunity, and antigen-presentation signaling pathways in high CD19 TIB MIBC patients. Moreover, CD19 TIB co-localized with activated CD4 TIT and expressed surface markers characteristic of antigen-presenting cells. Finally, an antigen-presentation assay demonstrated the antigen-presentation function of CD19 TIB.

Conclusion: CD19 TIB was identified as an independent prognostic factor, which could predict for post-surgery survival and platinum-based ACT benefits in MIBC. CD19 TIB serve as antigen-presenting cells (APCs) to activate CD4 TIT in the tumor environment of MIBC.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1007/s00262-018-2250-9DOI Listing
January 2019

Febuxostat in the treatment of gout patients with low serum uric acid level: 1-year finding of efficacy and safety study.

Clin Rheumatol 2018 Nov 9;37(11):3107-3113. Epub 2018 Sep 9.

Department of Rheumatology and Immunology, Nanjing First Hospital, Nanjing Medical University, Nanjing, 210006, China.

To retrospectively analyze the efficacy and safety of febuxostat on gout patients with low serum uric acid level. A study was conducted in Nanjing First Hospital from October 2015 to September 2016. Thirty nine acute gouty arthritis patients from the emergency and outpatient department were included. Patients met the 2015 Gout Classification Criteria revised by American College of Rheumatology/European League Against Rheumatism (ACR/EULAR) and had urate deposition around the joints detected by dual-energy computerized tomography (DECT). Patients whose serum uric acid (SUA) were between 5.0 and 7.0 mg/dl (300-420 μmol/l) received febuxostat treatment to maintain the SUA level between 3.0 and 5.0 mg/dl for 1 year. Efficacy and safety of febuxostat were observed during the process. Three of 39 subjects were excluded because of adverse events (AEs) after receiving an initial febuxostat treatment for 2 months. Thirty six subjects were enrolled. The mean SUA level was reduced significantly from 6.51 ± 0.28 mg/dl at baseline to 4.24 ± 0.38 mg/dl and SUA of all subjects decreased by 34.8% compared with baseline. After 1-year treatment, the volume of tophus was reduced approximately 62.8%. Serum creatinine decreased stepwise in 8 gout patients with chronic kidney diseases from 162.5 ± 9.2 μmol/l to 131.4 ± 11.0 μmol/l. Two months after initiation of treatment, the number of gout flares began to markedly decrease and almost did not occur after 1 year. After the 1-year treatment of febuxostat, the average SUA level declined significantly, and the renal function improved gradually. There was nearly complete abolition of gout flares by the end of the study. Tophi resolved markedly compared with baseline as assessed by DECT. Furthermore, only a few people experienced adverse events. Febuxostat has a notable effect for gout patients in the lower SUA level range.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1007/s10067-018-4283-zDOI Listing
November 2018