Transcriptome-wide 1-methyladenosine functional profiling of messenger RNA and long non-coding RNA in bladder cancer.

Introduction: Post-transcriptional RNA modifications are crucial regulators of tumor development and progression. In many biological processes, N1-methyladenosine (m1A) plays a key role. However, little is known about the links between chemical modifications of messenger RNAs (mRNAs) and long noncoding RNAs (lncRNAs) and their function in bladder cancer (BLCA). Methods: Methylated RNA immunoprecipitation sequencing and RNA sequencing were performed to profile mRNA and lncRNA m1A methylation and expression in BLCA cells, with or without stable knockdown of the m1A methyltransferase tRNA methyltransferase 61A (TRMT61A). Results: The analysis of differentially methylated gene sites identified 16,941 peaks, 6,698 mRNAs, and 10,243 lncRNAs in the two groups. Gene ontology enrichment and Kyoto Encyclopedia of Genes and Genomes pathway analyses of the differentially methylated and expressed transcripts showed that m1A-regulated transcripts were mainly related to protein binding and signaling pathways in cancer. In addition, the differentially genes were identified that were also differentially m1A-modified and identified 14 mRNAs and 19 lncRNAs. Next, these mRNAs and lncRNAs were used to construct a lncRNA-microRNA-mRNA competing endogenous RNA network, which included 118 miRNAs, 15 lncRNAs, and 8 mRNAs. Finally, the m1A-modified transcripts, SCN2B and ENST00000536140, which are highly expressed in BLCA tissues, were associated with decreased overall patient survival. Discussion: This study revealed substantially different amounts and distributions of m1A in BLCA after TRMT61A knockdown and predicted cellular functions in which m1A may be involved, providing evidence that implicates m1A mRNA and lncRNA epitranscriptomic regulation in BLCA tumorigenesis and progression.

Gene mutation detection of urinary sediment cells for NMIBC early diagnose and prediction of NMIBC relapse after surgery.

Early diagnose of bladder cancer could lead to good prognosis and high 5-year-survival rate. Among bladder cancer, about 75% patients with were nonmuscle-invasive bladder cancer (NMIBC). Patients were painful and easily get infected during bladder cancer diagnosis, which mainly depends on invasive cystoscopy and low-sensitivity urine exfoliation cytology. Meanwhile, relapse after surgery was also becoming the major problem for patients. Exploring noninvasive, high-sensitivity, and painless method is very important and meaningful for NMIBC treatment.Firstly, we found potential related gene mutation sites for NMIBC by searching COSMIC database and related study. Urinary sediment cells of patients both in normal group (patients with benign) and NMIMC group were collected before and after operation for potential gene mutation site detecting. Meanwhile, the urinary sediment cells of relapse patients and good prognosis people in NMIBC group after surgery were also collected for further Gene mutation detection and NMIBC relapse after surgery prediction.Fourteen genes (152 mutation sites) were selected between 95 NMIBC patients and 67 control patients, which were FGFR3, TP53, PIK3CA, and others. Compared with control group, mutation ratio of above 14 genes was higher in NMIBC group. NMIBC diagnose model was established by 5 times cross-validation and had a good effects, which included the all mutation site in FGFR3, TP53, PIK3CA, ARID1A, STAG2, and KTM2D. On the contrary, the relapse rate was 30.5% among 95 patients for about 1.5-year follow-up time. Compared with control group, smoking rate and tumor grade were higher in relapse group. Meanwhile, mutation rate of FGFR3, TP53, PIK3CA, ERBB3, and TSC1 in relapse group were higher than that in normal group. According to the mutation sites of FGFR3, TP53, PIK3CA, and ERBB3 and the combination of urinary sediment cells genetic mutation and relapse status, a predicted model for NMIBC relapse was also established, which had 90% accuracy.The diagnosed NMIBC model (based on FGFR3, TP53, PIK3CA, ARID1A, STAG2, and KTM2D gene mutation) and predicted relapse model (based on FGFR3, TP53, PIK3CA, and ERBB3 gene mutation) possess high accuracy and would be applied in early diagnose and early predicting relapse of patients.

Early Intratracheal Administration of Corticosteroid and Pulmonary Surfactant for Preventing Bronchopulmonary Dysplasia in Preterm Infants with Neonatal Respiratory Distress Syndrome: A Meta-analysis.

There is uncertain result with regard to the use of inhalation or instillation steroids to prevent bronchopulmonary dysplasia in preterm infants. This meta-analysis was designed to evaluate the efficacy and safety of early airway administration (within 2 days after birth) of corticosteroids and pulmonary surfactant (PS) for preventing bronchopulmonary dysplasia (BPD) in premature infants with neonatal respiratory distress syndrome (NRDS). The related studies were retrieved in PubMed, EMBASE, the Cochrane Library, Clinical Trial, CNKI, Wanfang and VIP Database from inception to August 2018. Two reviewers independently screened the studies to ensure that all patients with diagnosis of NRDS were enrolled to studies within 1 day after birth, assessed the quality of included studies by GRADEpro system and extracted the data for review. The meta-analysis was performed by RevMan 5.2 software. A subgroup analysis about inhaled corticosteroid (ICS) delivery method was made between ICS inhalation subgroup [inhalation of ICS by nebulizer or metered dose inhaler (MDI)] and ICS intratracheal instillation subgroup (PS used as a vehicle). Eight randomized controlled trials were enrolled in the meta-analysis, 5 trials of which stated the randomized method, grouping and blinded method, and the follow-up procedures were reported. GRADEpro system showed high quality of 4 trials (5 articles), and the rest 4 trials had moderate quality. Meta-analysis showed that the incidence of BPD was decreased in ICS group, the relative risk (RR) was 0.56 (95% CI: 0.42-0.76), and similar trends were found in ICS inhalation subgroup and ICS intratracheal instillation subgroup, with the corresponding RR being 0.58 (95% CI: 0.41-0.82) and 0.47 (95% CI: 0.24-0.95) respectively. ICS could also significantly reduce the mortality risk as compared with placebo control group (RR: 0.67; 95% CI: 0.45-0.99), with RR of ICS inhalation subgroup and ICS intratracheal instillation subgroup being 0.81 (95% CI: 0.34-1.94) and 0.64 (95% CI: 0.41-0.99) respectively. Moreover, the percentage of infants using PS more than one time was lower in ICS group than in the placebo control group, with the RR and 95% CI being 0.55 (95% CI: 0.45-0.67), and that in ICS intratracheal instillation subgroup lower than in ICS inhalation subgroup (RR: 0.56; 95% CI: 0.45-0.69, and RR: 0.35; 95% CI: 0.08-1.52 respectively). There was no significant difference in the incidence of infection or retinopathy of prematurity and neuro-motor system impairment between ICS group and placebo control group, with the corresponding RR being 0.95 (95% CI: 0.59-1.52), 0.92 (95% CI: 0.62-1.38) and 1.13 (95% CI: 0.92-1.39), respectively. It was concluded that early administration of ICS and PS is an effective and safe option for preterm infants with NRDS in preventing BPD and reducing mortality, decreasing the additional PS usage, especially for the ICS intratracheal instillation subgroup. Furthermore, the appropriate dose and duration of ICS, combined use of inhalation or instillation of ICS with PS and the long-term safety of airway administration of corticosteroids need to be assessed in large trials.

Expression of miRNA-630 in bladder urothelial carcinoma and its clinical significance.

Many studies informed that microRNAs (miRNAs) could function as diagnostic and prognostic indicators in several cancers. The aims of this study were to explore the expression of miR-630 in bladder urothelial carcinoma and its clinical significance for the evaluation of cancer prognosis. A total of 116 patients with bladder urothelial carcinoma were obtained in this retrospective study between May, 2012 and Sep. 2015. Quantitative real-time PCR (qRT-PCR) was conducted to evaluate the expression level of miR-630. The chi-square test was used to examine the associations between miR-630 expression and the clinicopathological features. The Kaplan-Meier method was conducted to explore the survival status of urothelial carcinoma patients. The log-rank test was used to analyze differences in survival rate. The results showed an obvious increase in miR-630 expression from normal bladder to bladder urothelial carcinoma (P=0.027). Additionally, patients with higher miR-630 expression had significantly shorter disease-free survival (DFS) (P=0.043) and overall survival (OS) (P=0.038) than those with lower miR-630 expression. Furthermore, multivariate analysis revealed that up-regulation of miR-630 was an independent prognostic factor for both DFS (P=0.042) and OS (P=0.046). It was demonstrated that miR-630 may be a novel and valuable prognostic factor for bladder urothelial carcinoma.

Expression and significance of m1A transmethylase, hTrm6p/hTrm61p and its related gene hTrm6/hTrm61 in bladder urothelial carcinoma.

OBJECTIVE: To discuss the expression of hTrm6p/hTrm61p in bladder urothelial carcinoma tissue and its relationship with m1A level in urine, as well as the influences of hTrm6/hTrm61 on the proliferation and apoptosis of cancer cell line on urothelium. METHODS: m1A levels in urine of 32 patients of bladder urothelial carcinoma and normal people were detected by HPLC/ESI-Q-TOF-MS, hTrm6p/hTrm61p expression levels in cancer tissue and para-carcinoma tissue of the same patient were detected by western blotting, and hTrm61 expressions of cancer cell line T24, 5637, and EJ of bladder urothelium and kidney cell line HEK-293 of human embryo were detected by RT-qPCR. The hTrm61 high-expression cell line is selected to detect the situation of proliferation and apoptosis by CCK-8 and flow cytometry (FCM) after knocking out hTrm61 with siRNA. RESULTS: m1A level in urine of carcinoma of urinary bladder is significantly higher than that of normal people, and hTrm6p/hTrm61p expression level in cancer tissue is significantly higher than that in para-carcinoma tissue and has linear correlation with m1A level in urine. The hTrm61 expression in cell line 5637 is significantly higher than that of T24, EJ, and HEK293, and apoptosis is significantly affected after hTrm61 is knocked out from 5637 cell line. CONCLUSION: High-level expression of hTrm6p/hTrm61p is an important reason for high emission of m1A in urine, and hTrm6/hTrm61 promotes the occurrence of carcinoma of urinary bladder.

Lenalidomide in treating patients with castration-resistant prostate cancer.

BACKGROUND: This analysis was conducted to evaluate the efficacy and safety of lenalidomide based regimen in treating patients with castration-resistant prostate cancer. MATERIALS AND METHODS: Clinical studies evaluating the efficacy and safety of lenalidomide based regimens on response and safety for patients with castration-resistant prostate cancer were identified using a predefined search strategy. A pooled response rate (rate of PSA level decline of ≥50%) to treatment was calculated. RESULTS: In lenalidomide based regimen, 3 clinical studies which including 98 patients with castration-resistant prostate cancer were considered eligible for inclusion. These lenalidomide based regimens included cisplatin, doxorubicin, or GM-CSF. Pooled analysis suggested that, in all patients, the pooled PSA level decline of ≥50% was 13.3% (13/98) in lenalidomide based regimens. Fatigue, nausea and vomitting were the main side effects. No grade III or IV renal or liver toxicity were observed. No treatment related death occurred in patients with lenalidomide based regimens. CONCLUSIONS: This evidence based analysis suggests that lenalidomide based regimens are associated with mild response rate and acceptable toxicities for treating patients with castration-resistant prostate cancer.

Urinary modified nucleosides as novel biomarkers for diagnosis and prognostic monitoring of urothelial bladder cancer.

AIMS AND BACKGROUND: To ascertain the value of the detection of urinary modified nucleosides in the early diagnosis and prognostic monitoring of urothelial bladder cancer. METHODS: One hundred seventeen patients with urothelial bladder carcinoma and 66 healthy volunteers were included in the study. High-performance liquid chromatography/electrospray ionization quadrupole time-of-flight mass spectrometry (HPLC/ESI-Q-TOF-MS) was used to measure the levels of urinary modified nucleosides in the bladder cancer and control groups. Postoperative monitoring was done every 3 months in patients with noninvasive carcinoma; 85 patients attended the 1-year follow-up visit. RESULTS: The levels of m1A, ac4C, O6-MeG and 1-MeI were significantly higher in cases than controls (P < 0.05). The highest sensitivity (92.45%) and specificity (87.50%) were obtained when 1-MeI detection was combined with m1A detection. The m1A and 1-MeI levels 3 months after operation in both patient groups were significantly lower than the preoperative levels (P < 0.01). The no-recurrence group subsequently maintained low levels, but in the recurrence group the levels rose again almost to preoperative values. At 6, 9 and 12 months after operation, the m1A and 1-MeI levels of the recurrence group were higher than those of the no-recurrence group and the control group (P < 0.01). CONCLUSIONS: Urinary modified nucleosides might become novel tumor markers that will facilitate the clinical management and will be helpful in the diagnosis and follow-up of urothelial bladder cancer. m1A and 1-MeI appear to be most promising for clinical use and be worthy of further study in the near future.

Urodynamic parameters development and complications of clean intermittent self-catheterization in Chinese schoolchildren with neurogenic underactive bladder.

OBJECTIVE: To evaluate the urodynamic parameters, development of bladder function and complications of clean intermittent self-catheterization (CIC) in Chinese schoolchildren with neurogenic underactive bladder. METHODS: Ninety-three children with neurogenic underactive bladder were successfully treated with CIC or combined with oxybutynin for two years follow-up. According to bladder compliance before CIC, they were subdivided into a normal bladder compliance (NBC) group and a low bladder compliance (LBC) group. Urodynamic parameters and complications were recorded. RESULTS: At follow-up, the incidence of neurogenic detrusor overactivity was found to have significantly decreased in both groups. Moreover, maximum cystometric capacity (CC) and relatively safe CC in the NBC group was significantly higher than those before CIC. However, relatively safe CC was significantly lower than that before CIC, and detrusor leakage point pressure was significantly higher than that before CIC in the LBC group. The incidences of bacteriuria, vesicureteral reflux (VUR), febrile urinary tract infections (UTI) and macroscopic hematuria were, respectively, 62, 13, 25 and 15%, and those of VUR and febrile UTI in the LBC group were significantly higher than those in the NBC group. CONCLUSION: For these cases, the complications of CIC are rare, and bladder compliance seems to be correlated with the development of bladder function and complications during CIC.

[Transperitoneal laparoscopic adrenalectomy for adrenal neoplasm: a report of 371 cases].

BACKGROUND AND OBJECTIVE: Open adrenalectomy has been almost replaced by mini-invasive laparoscopic surgery. There are two popular mini-invasive laparoscopic adrenalectomy approaches: retroperitoneal and transperitoneal approaches. This study was to summarize our experience in transperitoneal laparoscopic adrenalectomy. METHODS: In total 371 cases undergoing transperitoneal adrenalectomy in the First Affiliated Hospital of Zhengzhou University from February 2003 to August 2008 were reviewed retrospectively. There were 127 cases of primary hyperaldosteronism adenoma, 117 cases of Cushing's adenoma, 58 cases of phaeochromo-cytoma, 37 cases of incidentoma and 32 cases of other types. The type of adrenal diseases, operating time, blood loss, complications and prognosis were summarized and the operating method was analyzed. RESULTS: Three hundred and sixty-five out of 371 patients (98.4%) were successfully operated, five cases (1.4%) were transferred to open surgery, and one patient gave up surgery due to extensive invasion. The operating time was 40-240 min (average, 70 min). The blood loss was 20-1000 ml (average, 80 ml). Two patients suffered from diaphragm injuries, one patient had right renal vein injury and one had colon injury. The mean time of hospital stay was five days. CONCLUSION: Transperitoneal laparoscopic adrenalectomy is one of the favorable approaches for the treatment of adrenal neoplasm.

Association of NAT2, GSTM1, GSTT1, CYP2A6, and CYP2A13 gene polymorphisms with susceptibility and clinicopathologic characteristics of bladder cancer in Central China.

OBJECTIVE: To explore the association of polymorphisms in N-acetyltransferase 2 (NAT2), glutathione S-transferase (GST), cytochrome P450 (CYP) 2A6, and CYP 2A13 genes with susceptibility and clinicopathologic characteristics of bladder cancer in a Chinese population. METHODS: In a hospital-based case-control study of 208 cases and 212 controls matched on age and gender, genotypes were determined by PCR-based methods. Risks were evaluated by unconditional logistic regression analysis. RESULTS: It was found that significant associations of the NAT2 slow-acetylator genotype (odds ratio, OR: 2.42; 95% confidence interval, CI: 1.47-3.99), GSTM1 null genotype (OR: 1.64; 95% CI: 1.11-2.42) and GSTM1/GSTT1-double null genotype (OR: 1.72; 95% CI: 1.00-2.95) with increased risk of bladder cancer. Conversely, carriers with at least one CYP2A6*4 allele showed lower risk than the non-carriers (OR: 0.47; 95% CI: 0.28-0.79). The adjusted ORs (95% CI) for smokers with NAT2 slow-acetylator, GSTM1 null, GSTM1/GSTT1-double null genotype, and variant CYP2A6 genotypes were 2.99 (1.44-6.25), 1.98 (1.13-3.48), 2.66 (1.22-5.81) and 0.41 (0.20-0.86), respectively. Furthermore, NAT2 slow-acetylator, GSTM1 null, and GSTM1/GSTT1-double null genotypes were associated with higher tumor grade (P=0.001, 0.022, and 0.036, respectively), and only NAT2 slow-acetylator genotype was associated with higher tumor stage (P=0.007). CYP2A13 was not associated with risk or tumor characteristics. CONCLUSION: It is suggested that NAT2 slow-acetylator, GSTM1 null, GSTM1/GSTT1-double null, and variant CYP2A6 genotypes may play important roles in the development of bladder cancer in Henan area, China.

[Association between genetic polymorphism of UGT1A7 and susceptibility of bladder cancer].

OBJECTIVE: To investigate the association between genetic polymorphism of UGT1A7 and susceptibility of bladder cancer. METHODS: Based upon a case-control study, UGT1A7 polymorphisms were determined by the semi-nested polymerase chain reaction (SN-PCR) and allele-specific polymerase chain reaction (AS-PCR) in 208 cases with bladder cancer and 205 non-tumor controls. Risks were evaluated by unconditional logistic regression analysis. RESULTS: The frequency of variant homozygous genotype in cases (20.7%) was higher than that in controls (12.2%) and the difference was statistically significant [P < 0.05, OR = 2.16 (1.18 - 3.96)]. The frequency of variant allele (*)3 in cases was higher than that in controls (27.9%, 20.5% respectively) and the difference was statistically significant [P = 0.009, OR = 1.56 (95%CI: 1.12 - 2.18)]. The smokers with variant homozygous and heterozygous genotypes showed an increased risk of bladder cancer compared with those with wild genotype [2.16 (95%CI: 1.07 - 4.36), 2.64 (95%CI: 1.02 - 6.80) respectively]. There was no association between the UGT1A7 polymorphisms and the pathological grade and clinical stage of bladder cancer (both P > 0.05). CONCLUSION: The genetic polymorphisms of UGT1A7 are associated with the susceptibility of bladder cancer and have interactions with smoking in bladder carcinogenesis.

Bed-wetting in Chinese children: epidemiology and predictive factors.

AIMS: To estimate the prevalence and severity of bed-wetting in 1-18-year-old Chinese children. MATERIALS AND METHODS: A cross-sectional study of bed-wetting was performed by using 13,515 self-administered questionnaires distributed to the parents of 1-18-year-old Chinese children in Henan province. The prevalence of bed-wetting was determined. The relationship of wetting to age, gender, community characteristics (rural or urban), arousal dysfunction, associated day-time symptoms (frequency, urgency, and incontinence), and family history were analyzed. RESULTS: There was a response rate of 87% (5,978 boys and 5,786 girls). The overall prevalence of bed-wetting was 23.03% in those aged 1-4, 5.66% in those 5-12, and 1.37% in those 13-18. When a logistic regression analysis was applied to determine risk factors for the bed-wetting, a positive relationship was seen with male gender and living in rural areas. Further, living in rural areas, arousal dysfunction, and associated day symptoms were significantly related to more severe bed-wetting. Only 3.64% of the children had undergone professional evaluation. CONCLUSION: The prevalence of bed-wetting is significant in Chinese children, but lower than in most western countries, which is likely due to cultural differences. Living in rural areas, having arousal dysfunction, and having associated day-time symptoms may be predicative factors for marked bed-wetting.

Efficient screening method of the thiopurine methyltransferase polymorphisms for patients considering taking thiopurine drugs in a Chinese Han population in Henan Province (central China).

BACKGROUND: Thiopurine S-methyltransferase (TPMT) is an enzyme that catalyzed the S-methylation of thiopurine drugs. TPMT activity exhibits an interindividual variability, mainly as a result of genetic polymorphism. Patients with intermediate or deficient TPMT activity are at risk for toxicity after receiving standard doses of thiopurine drugs. We determined a cut-off concentration of the TPMT activity assay less than which genotyping of the TPMT gene should be performed. In addition, the influence of hemodialysis on TPMT activity in uremic patients was examined. METHODS: In 248 healthy subjects and 30 uremic patients, PCR-based methods were used to analyze the most common functional mutations TPMT2, 3A, 3B and 3C. A HPLC assay was used to measure erythrocyte TPMT activity in the whole population. RESULTS: Seven TPMT3C heterozygotes were identified, while TPMT2, 3A and 3B alleles were not detected in 248 healthy subjects. The frequency of TPMT3C allele was 1.4% (7/496). The TPMT activity in healthy subjects was normally distributed, ranged from 6.09 to 28.65 nmol/h/ml pRBC with a mean of 16.03 +/- 4.16 nmol/h/ml pRBC. The cut-off for high TPMT activity and intermediate TPMT activity was 10.07 nmol/h/ml pRBC. There were 19 intermediate activity healthy subjects (7.7%) and 229 high activity healthy subjects (92.3%), and no TPMT deficiency subject was found. All of the 229 healthy subjects with high activity had no mutant alleles, while 7 of the 19 subjects with intermediate activity had a mutant allele. Phenotypes were in good agreement with genotypes for 95% of subjects. The uremic patients were all homozygous for the wild-type allele whose TPMT activity was activated significantly before hemodialysis compared with TPMT activity after hemodialysis. CONCLUSIONS: We defined the cut-off values for the TPMT phenotyping assay at 10.07 nmol/h/ml pRBC, less than which additional genotyping elucidates the individual risk for drug therapy. In uremic patients, TPMT activity is increased by some uremic factors, and dialysis shifted their TPMT activity close to that of a healthy control group.

Is it possible to use urodynamic variables to predict upper urinary tract dilatation in children with neurogenic bladder-sphincter dysfunction?

OBJECTIVE: To investigate the possibility of using urodynamic variables to predict upper urinary tract dilatation (UUTD) in children with neurogenic bladder-sphincter dysfunction (NBSD). PATIENTS AND METHODS: The study included 200 children with NBSD, of whom 103 had UUTD and 97 did not; they were examined using routine urological, neurological and urodynamic methods. The group with UUTD was divided into three subgroups (group 1-3, from mild to severe hydronephrosis). A urodynamic risk score (URS) was calculated, including a detrusor leak-point pressure (DLPP) of >40 cmH2O, a bladder compliance (BC) of <9 mL/cmH2O and evidence of acontractile detrusor (ACD). RESULTS: The postvoid residual urine volume (PVR), DLPP, incidences of ACD and DLPP of >40 cmH2O were greater and the BC significantly less in groups 1-3 than in the control group. Moreover, the BC decreased, while the PVR, DLPP and the incidence of DLPP of >40 cmH2O were significantly higher in group 3 than in group 2. The relative safe cystometric capacity of groups 2 and 3 were lower, respectively, than that of the control and group 1, and the relative unsafe cystometric capacity (RUCC) and relative risk rate of cystometric capacity (RRRCC) were significantly greater with the severity of UUTD. The maximum detrusor pressure on voiding or at maximum flow rate, and the Abrams-Griffiths number for voluntary contractile bladders, of the UUTD group were significantly higher than those of the control group. There was a positive correlation between URS and UUTD. CONCLUSIONS: The selective use of urodynamic variables might be valuable for predicting the risk of UUTD in children with NBSD. Decreased BC, and increased DLPP and ACD are the main urodynamic risk factors, and they reciprocally increase the occurrence and grades of UUTD. The grades of UUTD are compatible with increases in RUCC, RRRCC and URS.