N6-methyladenosine (m6A) is the most prevalent reversible modification in eukaryotic mRNA, and it plays a critical role in tumor progression. The purpose of this study was to investigate the function and regulatory mechanisms of the methyltransferase METTL3 in renal cell carcinoma (RCC). METTL3 expression was upregulated and predicted a poor prognosis in patients with advanced RCC. METTL3 facilitated the proliferation, migration, and invasion of RCC cells, depending on its methylase activity. METTL3 positively regulated the expression of PLOD2, and both genes were triggered under prolonged hypoxia. Mechanistically, hypoxia-induced the binding of HIF-1α to the METTL3 promoter, which enhanced its transcriptional activity. METTL3-mediated m6A modifications of PLOD2 mRNA at 3’UTR region, promoting the translation of PLOD2 protein. Furthermore, silencing METTL3 impaired RCC progression in vitro. In vivo, administration of highly potent and selective METTL3 inhibitor STM2457 showed anti-tumor effects, whereas AAV9-mediated re-transduction of PLOD2 largely abolished the above phenomenon in a subcutaneous mouse model. These findings reveal that hypoxia and HIF-driven METTL3 transcription promote RCC progression by increasing PLOD2 expression in an m6A-dependent manner, suggesting that METTL3 may serve as a novel pharmaceutical intervention for RCC.

Tumors of the male genitourinary system are of great concern to the health of men worldwide. Although emerging experiment-based evidence indicates an association between hepcidin and such cancers, an integrated analysis is still lacking. For this reason, in this study, we determined the underlying oncogenic functions of hepcidin in common male genitourinary system tumors, including bladder urothelial carcinoma (BLCA), kidney chromophobe (KICH), kidney renal clear cell carcinoma (KIRC), kidney renal papillary cell carcinoma (KIRP), prostate adenocarcinoma (PRAD), and testicular germ cell tumors (TGCT) according to the data from The Cancer Genome Atlas. We found that hepcidin was highly expressed in kidney and testicular cancers. Meanwhile, the expression level of hepcidin was distinctly associated with the prognosis and immune cell infiltration in male patients with certain genitourinary system cancers, especially in KIRC. Elevated hepcidin levels also present as a risk factor in male genitourinary system tumors. Moreover, enrichment analyses revealed that some of the principal associated signaling pathways involving hepcidin and its related genes are identified as tumorigenesis-related. Immunofluorescence staining confirmed the conclusion of our immune infiltration analysis in KIRC tissue. In this study, for the first time, we provided evidence for the oncogenic function of hepcidin in different types of male genitourinary system tumors.

This study aimed to evaluate the value of cystatin C (Cys C) in predicting the perioperative and long-term prognosis of renal transplantation (RT). The clinical data of 198 RT recipients were collected. Blood samples were obtained daily until 7 d after transplantation and then discharge day to determine the serum levels of Cys C. The receiver-operating characteristic (ROC) analysis and the area under the curve (AUC) were used to determine the diagnostic accuracy of Cys C for delayed graft function (DGF). The presence of shrunken pore syndrome (SPS) with a cystatin C-based estimate of glomerular filtration rate less than 70% of a creatinine-based estimate, was also evaluated as a prognostic factor for the development of DGF. The serum Cys C levels of patients with DGF were higher than those of the non-DGF group. Cys C showed a higher AUC (0.928) in the ROC analysis than did sCr (0.862). Compared to the non-SPS group, there were more patients diagnosed with SPS in the DGF group (p < .05). The follow-up data showed that patients diagnosed with SPS had higher levels of sCr and Cys C compared to other patients, suggesting a poor long-term prognosis. Our findings suggest that Cys C is a sensitive indicator of renal function during the perioperative period. Cys C at a concentration of 4.9 mg/L had the highest sum of sensitivity and specificity for prediction of DGF, with a sensitivity of 0.889 and a specificity of 0.8. SPS is associated with the development of DGF and the poor long-term prognosis of RT.

Abstract Background Bladder cancer (BC) is a common type of cancer that involves tumors of the urinary system and poses a serious threat to human health. Long noncoding RNAs (lncRNAs) have emerged as crucial biomarkers and regulators in many cancers. Novel lncRNA biomarkers in BC urgently need to be investigated in regard to its function and regulatory mechanisms. Methods Identification of differentially expressed lncRNAs in BC tissue was performed via microarray analysis. To investigate the biological functions of LINC00612, loss-of-function and gain-of-function experiments were performed in vitro and in vivo. Bioinformatics analysis, dual-luciferase reporter assays, AGO2-RIP assays, RNA pull-down assays, real-time quantitative PCR (RT-qPCR) arrays, fluorescence in situ hybridization assays, and western blot assays were conducted to explore the underlying mechanisms of competitive endogenous RNAs (ceRNAs). Results LINC00612 was upregulated in BC tissues and cell lines. Functionally, downregulation of LINC00612 inhibited cell proliferation and invasion in vitro and in vivo, whereas overexpression of LINC00612 resulted in the opposite effects. Bioinformatics analysis and luciferase assays revealed that miR-590 was a direct target of LINC0061, which was validated by dual-luciferase reporter assays, AGO2-RIP assays, RNA pull-down assays, RT-qPCR arrays, and rescue experiments. Additionally, miR-590 was shown to directly target the PHD finger protein 14 (PHF14) gene. LNIC00612 modulated the expression of E-cadherin and vimentin by competitively sponging miR-590 to elevate the expression of PHF14, thus affecting BC cellular epithelial-mesenchymal transition (EMT). Conclusions Our results indicate that LINC00612 enhances the proliferation and invasion ability of BC cells by sponging miR-590 to upregulate PHF14 expression and promote BC cellular EMT, suggesting that LINC00612 may act as a potential biomarker and therapeutic target for BC.

肾移植是终末期肾病患者的首选治疗方法。移植的主要目标是通过免疫耐受减轻急、慢性排斥反应，延长供肾生存时间。大鼠肾移植模型是研究免疫排斥过程和评估免疫耐受等基础研究的重要工具 [1]。自从1965年首次建立大鼠肾移植模型以来，国内外学者对其实验技术不断地进行创新和改进，以期建立成功率更高、更易稳定存活的大鼠肾移植实验模型。在大鼠身上实施肾移植需要广泛的外科训练和专业知识，才能达到较高的存活率。大鼠肾移植模型的成功关键在于供受体的动静脉血管吻合，有报道运用套管法、内套法等各种血管吻合的方法。我们应用临床上的一次性静脉留置针辅助供受体的血管吻合，构建简单、省时、易于掌握的大鼠肾移植模型，现将方法报道如下。

Short-chain fatty acids (SCFAs), the end products of fermentation carried out by the intestinal microbiota, were demonstrated to produce anti-oxidant and anti-inflammatory effects. Butyrate, part of the SCFAs, also shows the same effect. Renal ischemia/reperfusion (I/R) injury commonly occurs in renal transplantation and is often accompanied by oxidative stresses and inflammatory responses. In this study, we explore butyrate effect on renal I/R injury and SCFAs changes in renal transplant. Male Sprague-Dawley rats were pretreated with butyrate as research, and underwent the surgery of renal ischemia for 45 min followed by reperfusion. 90 rats were randomly divided into 3 groups (n=30 each group): (1) sham-operated group; (2) butyrate-treated group; (3) control group. The samples of blood and renal were collected immediately for further studies. Thirty-two patients were enrolled to investigate the levels of SCFAs after the renal transplantation. Rats model showed that butyrate treatments significantly enhanced the function and structure of kidney, as evidenced by the lower serum creatinine levels and less pathological damages of renal tissue. With the recovery of renal function after renal transplantation, SCFAs increased, which were negatively correlated with creatinine. Butyrate expressed like SCFAs. In this study, we demonstrated that butyrate increased with the recovery of renal function after renal transplantation. Most importantly, butyrate treatments alleviated the renal damages caused by I/R via the upregulation of intracellular oxidant stress and inflammations.

BACKGROUND:It has been widely recognized that exosomal miRNAs can participate in the pathogenesis of different renal disorders and serve as disease biomarkers. Although kidney biopsy is still the gold standard for diagnosing and monitoring immunoglobulin A nephropathy (IgAN), it is highly required to identify new and effective noninvasive biomarkers for IgAN, the most frequently detected primary glomerulonephritis worldwide.METHODS:Plasma and urinary exosomes were extracted by PEG precipitation. Size and morphological characteristics of plasma and urinary exosomes were observed by transmission electron microscopy and nanoparticle tracking analysis. The levels of plasma and urinary exosomes were revealed by Western blotting. The expressions of target miRNAs were revealed by in situ hybridization and qRT-PCR.RESULTS:The levels of plasma and urinary exosomes were remarkably enhanced in IgAN patients compared with healthy controls (HCs). The expressions of miR-4639 and miR-210 in IgAN patients were significantly higher in contrast to the individuals with membranous nephropathy, minimal change nephrosis, diabetic nephropathy, or HC. These also played a valuable role in assessing the kidney function and the level of proteinuria. Furthermore, plasma and urinary exosomal miR-4639 expression was associated with more serious and active histological activity (mesangial hypercellularity, crescent, and C3 complement deposition). With an average follow-up of 8 months, miR-4639 and miR-210 expressions in plasma and urinary exosomes were higher in patients with progressive IgAN. Plasma exosomal miR-4639 and miR-210 were better than proteinuria (g/24 h) to estimate renal outcomes.CONCLUSION:Exosomal miR-4639 and miR-210 could be used as valid biomarkers to assist in diagnosis, evaluate severity, and assess disease development of IgAN.

本研究旨在探讨丁酸对肾缺血再灌注损伤的影响，现报道如下。

The present study examined the expression levels of ferroportin, a transmembrane protein that transports iron from the inside of a cell to the outside, in the prostate cancer PC3, DU145 and LNCAP cell lines, in the normal prostate RWPE2 cell line, and in tissue samples from different differentiation stages of prostatic carcinoma and prostatic hyperplasia. The study also investigated the role of ferroportin protein expression in the diagnosis and prognosis of prostate cancer. Reverse transcription-quantitative polymerase chain reaction and western blot analysis were employed to measure the mRNA and protein expression levels of ferroportin in the PC3, DU145, LNCAP and RWPE2 cells. Immunohistochemistry was used to determine ferroportin protein expression in the prostate cancer and prostatic hyperplasia tissues. Compared with the normal prostate RWPE2 cells, ferroportin protein expression was significantly lower in the prostate cancer PC3, DU145 and LNCAP cells (P<0.05). Compared with the prostatic hyperplasia tissues, ferroportin protein expression was significantly reduced in the prostate cancer tissues (P<0.05). Overall, the expression levels of ferroportin in the prostate cancer tissues were lower than those in the normal prostate tissues, which may provide valuable clinical information for the diagnosis and prediction of disease progression in prostate cancer, and may indicate a potential therapeutic target for treating prostate cancer by regulating iron metabolism.

Prostate cancer (PCa) is the most commonly diagnosed cancer in males and the fifth most common cause of cancer death worldwide. Previous studies indicated that miR-18a-5p modulated epithelial-mesenchymal transition in breast cancer via targeting SREBP1 forming a co-repressor complex with Snail and HDAC1/2. However, the function of miR-18a-5p in prostate cancer remains largely unknown. In this study, we identified miR-18a-5p as a tumor promoter in prostate cancer. miR-18a-5p expression was found upregulated in human prostate cancer tissues while SLC40A1 was down-regulated. Cell proliferation assay demonstrated that miR-18a-5p promoted prostate cancer cell proliferation. We also found SLC40A1 was downregulated by miR-18a-5p in prostate cancer cell lines. Restoration of SLC40A1 reversed the effects of miR-18a-5p in prostate cancer cells. Taken together, our results suggest that miR-18a-5p might function as a tumor-promoting factor in PCa and might contribute to its proliferation.

目的:通过对临床上发现的新型冠状病毒(2019-nCoV)IgG抗体阳性的病例进行分析,探讨血清学检测在辅助诊断和临床鉴别中的应用.方法:回顾性研究2020年3月19日～5月9日我院在7000余例筛查人群中检测发现新型冠状病毒IgG抗体阳性的160例病例,结合临床资料,与新型冠状病毒确诊患者和普通人群进行比较,分析IgG抗体阳性的原因和影响因素.结果:新型冠状病毒IgG抗体检测阳性率约为2.3％.总结临床资料发现IgG抗体阳性病例中73.1％(117/160)为肿瘤术后放化疗治疗的患者,92.5％(148/160)胸部CT平扫提示有慢性支气管炎,肺部纤维灶,肺部结节等症状.IgG抗体阳性病例白细胞、淋巴细胞、嗜酸性粒细胞较新型冠状病毒确诊患者表达高,具有统计学差异(P<0.05);IgG抗体阳性病例谷氨酰转肽酶,碱性磷酸酶,肌酐较新型冠状病毒确诊患者表达低,具有统计学差异(P<0.05);IgG抗体阳性病例与普通人群比较各项指标均无统计学差异(P>0.05).IgG抗体阳性病例结合鉴别诊断,流行病学史,接触感染史等均未发现有新型冠状病毒感染可能.结论:我院发现的新型冠状病毒IgG抗体阳性,结合临床资料均考虑是患者慢性感染、自身疾病、免疫等因素引起的假阳性.通过对IgG抗体阳性的评估,可以减少患者反复核酸检测,分析新型冠状病毒传播的可能,具有实际应用的意义.

A case of varicella pneumonia, hepatitis and pancreatitis after kidney transplantation was retrospectively analyzed. One week after kidney transplantation, the patient had a papule with pruritus, which was diagnosed as varicella by dermatologist as well as high-throughput sequencing. The patient was found to have pneumonia, hepatitis and pancreatitis. The individualized treatment regimen was used, including the dosage reduction of immunosuppressive agents, the blood drug concentration monitoring, antiviral therapy, anti-infection therapy, supportive treatment, and symptomatic alleviation for complications. The treatment was adjusted according to the indicators'variation. The timely review of the indicators and immunosuppressant blood concentration were performed to protect the transplanted kidney function, and the patient recovered in time. This rare case of postoperative complications of kidney transplantation were summarized and analyzed in order to accumulate clinical experience for the treatment of renal transplantation.

目的 了解新型冠状病毒(SARS-CoV-2)IgG抗体阳性患者的血常规及肾功能参数特点,鉴别诊断SARS-CoV-2感染患者及抗体阳性患者的血液指标,为早期临床诊断提供依据.方法 回顾性分析常州市第一人民医院2020年3月19-31日门诊检测SARS-CoV-2 IgG和IgM抗体阳性患者,体检中心健康成年人及传染病院核酸阳性确诊的SARS-CoV-2感染患者临床资料.所有患者均检测血常规、肾功能,以及影像学CT平扫胸部检查,选择有鉴别意义的标志物.结果 新型冠状病毒肺炎患者白细胞计数、淋巴细胞计数及嗜酸性粒细胞计数均明显低于健康成年人,差异均有统计学意义(P<0.05);肾功能参数中,新型冠状病毒肺炎患者的肌酐明显高于健康成年人,差异有统计学意义(P<0.05).SARS-CoV-2 IgM和IgG抗体阳性患者与健康成年人比较,白细胞计数差异无统计学意义(P>0.05);但淋巴细胞计数明显低于健康成年人,差异有统计学意义(P<0.05);肾功能参数中,两组肌酐水平比较,差异无统计学学意义(P>0.05).结论 SARS-CoV-2 IgM和Ig G抗体阳性提示为感染中后期或既往感染,抗体阳性核酸检测阴性是否为假阳性或无症状感染者自愈值得临床进一步观察研究.血常规及肾功能参数在鉴别诊断中存在一定意义,可为核酸及CT诊断提供补充.

目的:通过报道1例精索脂肪肉瘤的临床资料和分析基因检测罕见病的方法，并经过诊断分析和提供治疗方案，探讨其在临床上的应用价值和研究意义。方法:运用二代高通量基因测序技术，选取病理诊断为精索脂肪肉瘤的石蜡包埋组织，对与免疫治疗、靶向用药、肿瘤驱动和遗传性肿瘤相关基因的全部外显子及其部分内含子区域进行捕获测序，在DNA水平上分析疾病相关基因的突变形式，从而提供个体化的诊疗方案。结果:精索脂肪肉瘤驱动基因相关突变检测显示：CDK4扩增6.61倍，MDM2基因扩增6.61倍；免疫治疗相关检测显示：突变负荷(TMB)为0.56，微卫星稳定分析为稳定型；胚系突变检测未测出与遗传性肿瘤明确相关的致病突变；靶向药物检测显示：可能对哌柏西利敏感。结论:通过肿瘤基因检测对精索脂肪肉瘤的病例分析，可以对疾病的分子生物学机制、诊断方法和治疗有更深入的了解。对一些罕见病和特殊疾病，在临床上运用基因检测的方法，能指导医生认识疾病和进行研究。

目的 分析肾肉瘤样癌(SRCC)患者的临床表现,探讨基因检测在SRCC治疗中的作用.方法 回顾性分析2例SRCC患者的临床资料,并根据病理组织的高通量测序基因检测结果指导临床用药,为患者提供个体化的诊疗方案.术后持续随访观察.结果 2例SRCC患者均通过免疫组化明确病理诊断,术后随访发现病例1原位复发,病例2远处转移.基因检测提示原位复发对m T O R抑制剂依维莫司敏感;远处转移对PD1治疗敏感.原位复发患者予以依维莫司口服治疗,手术1年后仍在随访中;远处转移患者予以帕博利珠单抗治疗,半年后因多器官功能衰竭死亡.结论 基因检测可以为SRCC患者提供合适的治疗方案.

The purpose of the present study was to investigate the expressions of hsa-let-7c-5p and TGF- β signaling-related molecules and their correlations with clinical characteristics in chronic kidney disease (CKD). Twenty-three biopsy specimens of CKD patients and 20 negative control tissues were selected. Quantitative real-time PCR (qPCR) was used for the detection of hsa-let-7c-5p, transforming growth factor β (TGF- β ) and TGF- β receptor type 1 (TGF- β R1) expression levels. Target gene of hsa-let-7c-5p was verified by dual-luciferase reporter assay. A significant decrease of hsa-let-7c-5p expression in CKD tissue was found, compared with that of normal renal tissues (p<0.01). Expression levels of TGF- β in CKD were increased, compared with that of normal kidney tissue (p<0.001). The difference in the expression of TGF- β R1 between CKD tissues and normal renal tissues was not significant (p>0.05). A negative correlation was found between the expression of TGF- β and renal tissue hsa-let-7c-5p levels. Furthermore, hsa-let-7c-5p was identified to regulate TGF- β 1 by directly binding with the 167-173 site in the 3′ untranslated region. Decreased hsa-let-7c-5p levels in CKD patients was found to be associated with disease severity, which shows a negative correlation with proteinuria and creatinine levels, and a positive correlation with estimated glomerular filtration rate (eGFR), while relative TGF- β 1 expression had a positive correlation with creatinine level. In summary, changes in hsa-let-7c-5p expression and its target gene TGF- β are associated with the disease status of CKD. Let-7c-5p may contribute to the pathogenesis of renal fibrosis through TGF- β signaling, a potential diagnostic and therapeutic target of the disease.

Background: The aim of this study was to assess the diagnostic utility of iron homeostasis determinations for prediction of severity of COVID-19. Material/Methods: This was a retrospective study enrolling a total of 50 patients diagnosed with the novel coronavirus disease-19 (COVID-19) from February 27, 2020 to March 30, 2020, including a severe group (12 patients) and a mild group (38 patients). For the control group, 50 healthy people were examined during the same period. We compared clinical laboratory data and iron homeostasis biomarkers among the 3 groups. ROC curve analysis was used to assess diagnoses. Results: Patients diagnosed with severe COVID-19 had higher hepcidin and serum ferritin levels than in other groups (p<0.001). A combination test of hepcidin and serum ferritin provided the best specificity and sensitivity in the prognosis of COVID-19 severity. Logistic regression analysis showed hepcidin and serum ferritin independently contributed to the severity of COVID-19. Hepcidin and serum ferritin tandem testing predicted COVID-19 severity with 94.6% specificity, while hepcidin and serum ferritin parallel testing had a sensitivity of 95.7%. Conclusions: Iron homeostasis had a robust association with the occurrence of severe COVID-19. Iron homeostasis determinations were specific and sensitive for the early prediction of disease severity in COVID-19 patients and thus have clinical utility.

Aim: To systematically profile the global m 6 A modification pattern in clear cell renal cell carcinoma (ccRCC). Methods: m 6 A modification patterns in ccRCC and normal tissues were described via m 6 A sequencing and RNA sequencing, followed by bioinformatics analysis. m 6 A-related RNAs were immunoprecipitated and validated by quantitative real-time PCR (qPCR). Results: In total, 6919 new m 6 A peaks appeared with the disappearance of 5020 peaks in ccRCC samples. The unique m 6 A-related genes in ccRCC were associated with cancer-related pathways. We identified differentially expressed mRNA transcripts with hyper-methylated or hypo-methylated m 6 A peaks in ccRCC. Conclusion: This study presented the first m 6 A transcriptome-wide map of human ccRCC, which may shed lights on possible mechanisms of m 6 A-mediated gene expression regulation.

Objective: The microRNA expression profile of plasma exosomes in prostate cancer (PCa) is of critical importance in the disease exploration. This study aimed to explore the clinical application of exosomal miRNAs as biomarkers for PCa. Methods: Exosome-like vesicles of PCa patients and healthy controls were purified by differential centrifugation. The purified vesicles within the ranges of 50 and 100 nm were classified as exosomes according to the results of transmission electron microscopy and Western blot. Both, in vitro and in vivo, validations were performed by small RNA sequencing, CCK8, RT-qPCR, flow cytometry, Western blot, transwell and immunofluorescent staining assays. Results: High-throughput sequencing identified that 94 miRNAs were differentially expressed in PCa patients in comparison with healthy controls (P<0.01; fold change >= 2). Among them, 64 miRNAs were upregulated, and 30 miRNAs were downregulated. In comparison to the healthy controls, the expression levels of miR-217 were significantly upregulated, while miR-23b-3p were significantly downregulated in the exosomes and serum collected from PCa patients. Both, in vitro and in vivo, studies revealed that exosomes secreted by PCa cells with up-regulated miR-217 levels promoted cell proliferation and invasion; meanwhile, the exosomes with up-regulated miR-23b-3p levels inhibited cell proliferation and invasion. The epithelial-mesenchymal transition process may have been involved in the above-mentioned regulation. Conclusion: This study identified the dysregulated expression of exosomal miRNAs in PCa patients, including miR-217 and miR-23b-3p, by validating their function on proliferation and invasion in PCa cells. This regulation may have been affected by the epithelial-mesenchymal transition process, suggesting that they can be used as potential targets in the diagnosis and treatment of PCa.