MicroRNA (miRNA) are a class of small,non-coding,measuring 21-25 nucleotides in length RNA,which regulates protein levels post-transcriptionally through binding to 3'-untranslated region of target mRNA.miRNA involves in many physiological and pathological processes.Studies demonstrate that miRNA presents not only intracellular,but also in circulation and body fluids,e.g.urine,tear,ascetic fluid,and amniotic fluid,and these miRNA have become a novel biomarker and potential target of treatment,especially the serum miRNA.This review summarizes the recent advances of serum miRNA.

A method for the determination of methylmercury ( MeHg ) and ethylmercury ( EtHg ) in aquatic products was developed using gas chromatography-mass spectrometry with stable isotope-labelled internal standard. After ultrasonication assisted hydrochloric acid extraction, MeHg and EtHg in samples were extracted into toluene under the presence of sodium chloride and then back-extracted into cysteine aqueous solution. The MeHg and EtHg were released from their complexes with cysteine by adding cupric ions, and then derived with sodium tetraphenylborate. Under the optimal chromatographic conditions, MeHgPh and EtHgPh, the resulting derivatives, were separated completely on a DB-5MS capillary column and detected by electron impact ionization mass spectrometry in the selective ion monitoring ( SIM) mode, and quantified by a stable isotope dilution method using the d3-methylmercury as internal standard. The calibration curves were linear in the range of 1-500 μg/L of MeHg and EtHg. Concentration of 0. 828 mg Hg/kg with relative standard deviation ( RSD ) of 3 . 2% ( n=6 ) was obtained for MeHg in GBW 10029 . This was in good agreement with the certified values of (0. 84±0. 03) mg Hg/kg. The average recoveries were 94%-101% and 81%-104% for MeHg and EtHg spiked in aquatic samples, with RSDs of 1. 9%-4. 7% and 3. 1%-8. 2%(n=6), respectively. The limits of detection (S/N=3) of the two targets were 0. 1-0. 3μg/kg. This method was sensitive, accurate and could meet the demand of the determination of methylmercury and ethylmercury in aquatic products.

Objective To investigate the normal waveform,lantency and amplitude of auditory event-related potentials (ERPs) of exogenous components in children at different ages,and summarize the developmental rules and clinical perspectives of exogenous ERPs according to the International Federation of Clinical Neurophysiology (IFCN) Recommended Standards in 2009.Methods One hundred and thirty-six children aged from 3 to 17 years old (admitted in the departments from healthcare,orthopedics,general surgery,and respiratory and digestive diseases,and those recovered from respiratory infection and gastrointestinal infection),and healthy adults (from undergraduates of Chongqing medical university) were recruited in this study from 2008 to 2010.They were divided into 4 groups in accordance with their ages,that is,3 ～5,＞ 5 ～7,＞ 7 ～9,and ＞ 9 ～17 years old groups,and adult group (22 ～29 years old).ERPs were recorded from the midline site (Cz) with classical auditory Oddball paradigm.Each subject was tested twice or more to determine waveform repeatability.The overlapped graphs of ERPs under target and standard stimuli were obtained in the potentiometer.The exogenous ERPs,including P1,N1,P2 and N2,lantency and amplitude were measured.Results The latencies of exogenous ERPs under target and standard stimuli were shortened in a non-uniform manner with the increasing of age.No matter under target or standard stimuli,over 95％ of mean latencies of exogenous ERPs were within the range of 2SD.In addition to N1 component,the amplitude of P1,P2 and N2 components induced by target stimulus were in a trend of increased first and then decreased with the increasing of age.Compared with the children,the amplitude was much lower in the adults and adolescents (P ＜ 0.05).The N2 component induced by target stimulus was absent among 95％ healthy adults.The Standard deviations of amplitudes of exogenous components were great in all aged groups,indicating significant individual differences.Conclusion The latencies of exogenous ERPs are shortened with the increasing of age.Target stimulus will produce more stable results than the standard stimulus,and the ERPs under target stimulus can reflect the primary auditory cortex function of temporal lobe,and are independent on patients' cooperation.

Anultra-performancehydrophilicinteractionliquidchromatography-triplequadrupolemass spectrometric ( UPLC-MS/MS) method was developed for the determination of tetrodotoxin ( TTX) in human urine and plasma. After the sample was cleaned-up and concentrated by immunoaffinity column, the separation of the TTX was carried out on an Acquity UPLC BEH amide column (100 mm×2. 1 mm, 1. 7 μm) with gradient elution using mobile phases of 0. 1% ( V/V) formic acid in water and acetonitrile. The analyte was detected by positive electrospray ionization mass spectrometry in the multiple reaction monitoring ( MRM) mode, and quantified by external solvent standard calibration. The measuring ranges of TTX in urine and plasma were 0. 05-400 μg/L. The average recoveries were 92%-95% and 91%-96% for TTX respectively spiked in urine and plasma with relative standard deviations of 3 . 3%-7 . 2% and 3 . 9%-7 . 8% ( n=5 ) . The limits of detection (LOD, S/N=3) and limits of quantitation (LOQ, S/N=10) of TTX were 0. 02 μg/L and 0. 05μg/L for urine and plasma, respectively. This method is suitable for the detection of TTX in urine and plasma for both forensic and clinical purposes.

Objective:To summarize the clinical features, genetic testing and treatment efficacy of 6 children with Dent disease.Methods:Six children diagnosed with Dent disease in Guangzhou Women and Children′s Medical Center from January 2014 to March 2019 were enrolled.Their medical history, clinical manifestations, laboratory results, genetic test results, and proteinuria level, calciuria level and renal function after medication were measured.Results:All patients were male, with the onset age ranged from 1 to 9 years old.They were followed up for 6 months to 4 years.All the children had low molecular weight proteinuria.Urine protein electrophoresis showed that the ratio of low molecular weight proteinuria in only 2 cases was more than 50%.Renal biopsy suggested that all cases were combined with glomerular lesions.Five cases had hypercalciuria.Under the microscope, there were 5 cases of hematuria.Two case had rickets, and there was no renal calcium deposition and hypophosphatemia.Five cases were detected with CLCN5 mutations, of which p. C160Yfs*49 and p. G523D were first reported.One case had an OCRL1 mutation.Patients were treated with Hydrochlorothiazide and angiotensin converting enzyme inhibitor (ACEI). The 24 h urinary calcium level after treatment was lower than that before treatment [0.40 (0.24, 0.43) mmol/kg vs.0.12 (0.11, 0.14) mmol/kg, U=2.00, P<0.01]. However, there was no significant decrease in the 24 h-urinary protein level before and after treatment [77.09 (62.41, 88.01) mg/kg vs.80.33 (66.03, 92.52) mg/kg, U=12.00, P>0.05]. Conclusions:Dent disease is mainly characterized by low molecular weight proteinuria, and some patients may not be associated with hypercalciuria.Gene tests help to identify the disease type.ACEI and Hydrochlorothiazide can reduce the urinary calcium level, but cannot improve the level of urinary protein.

ObjectiveTo detect the expressions of Nuclear factor-erythroid 2 p45-related factor 2 (NRF2) and multidrug resistance-associated protein 2 (MRP2), and investigate their significance in primary gallbladder carcinoma. MethodsImmunohistochemistry SP assay and image analysis were used to detect the expressions of NRF2 and MRP2 protein in 59 patients with primary gallbladder carcinoma. ResultsA highly positive expression rates of NRF2 and MRP2 were found (76.3% and 74. 6%, respectively) in primary gallbladder carcinoma. The expressions of NRF2 and MRP2 had a significantly correlation with metastases, Nevin staging, and differentiation (P＜0.05), but there was no statistical association with sex and age. The expression of NRF2 had a positive correlation with MRP2 (r=0. 589,P＜0.05). Conclusion Both NRF2 and MRP2 were overexpressed in primary gallbladder carcinoma and they may play a role in the development of primary gallbladder carcinoma.

The correlation between the anatomic site of spinal cord injury and real-time conditions of bladder and urethral function was assessed in order to provide a reasonable basis for the clinical treatment of neurogenic bladder. A total of 134 patients with spinal cord injuries (105 males, 29 females; averaged 34.1 years old) were involved in this retrospective analysis, including urodynamic evaluation, clinical examination and imaging for anatomical position, and Bors-Comarr classification. The associations between the levels of injury and urodynamic findings were analyzed. The results showed that mean follow-up duration was 16.7 months (range 8-27 months). Complete spinal cord injuries occurred in 21 cases, and incomplete spinal cord injuries in 113 cases. Of the 43 patients with upper motor neuron (UMN) injuries, hyperreflexia and (or) detrusor sphincter dyssynergia were demonstrated in 30 (69.8%), 31 (72.1%) suffered low bladder compliance (less than 12.5 mL/cmH(2)O), 28 (65.1%) had high detrusor leak point pressures (greater than 40 cmH(2)O), and 34 (79.1%) had residual urine. Of the 91 patients with lower motor neuron (LMN) injuries, areflexia occurred in 78 (85.7%), high compliance in 75 (82.4%), low leak point pressures in 80 (87.9%), and residual urine in 87 (95.6%), respectively. The associations between the anatomical site of spinal cord injury and urodynamic findings were ill defined. In patients with spinal cord injury, this study revealed a significant association between the level of injury and the type of voiding dysfunction. The anatomical site of spinal cord injury can not be predicted in real-time condition of bladder and urethral function. Management of neurogenic bladder in patients with spinal cord injury must be based on urodynamic findings rather than inferences from the neurologic evaluation.

Objective To analyze the long-term prognosis and prognostic factors of idiopathic collapsing focal segmental glomerulosclerosis (FSGS) and not otherwise specified FSGS in children. Methods The clinical, pathology and follow-up data of patients with idiopathic collapsing FSGS and not otherwise specified FSGS were analyzed retrospectively by Kaplan-Meier method, univariate and multivariate Cox regression analysis. Results A total of 64 patients (29 idiopathic collapsing FSGS and 35 not otherwise specified FSGS) were diagnosed by renal biopsy. The 4-year renal survival rate of idiopathic collapsing FSGS and not otherwise specified FSGS were 48.3%, 74.3% respectively. Univariate analysis revealed that the renal survival time were 25.41±3.28 months in idiopathic collapsing patients, and 35.53±2.73 months in not otherwise specified patients. The different is significant (χ2=4.07,P=0.044). Multivariate Cox regression analysis showed that poor treatment response (HR=5.92, P<0.05) and renal insufficiency at early stage (HR=2.45, P<0.05) were independent risk factors of prognosis. Conclusions Compared with patients with not otherwise specified FSGS, the renal survival time is shorter in idiopathic collapsing FSGS patients. Patients with renal insufficiency and poor response to treatment have poorer prognosis.

Objective To investigate the effect of periurethral injection of insulin-like growth factor(IGF)-Ⅰ on the expression of IGF-Ⅰ and IGF-Ⅱ mRNA during regeneration period following urethral sphincter muscle injury in female rats. Methods Model of urethral sphincter muscle injury was made in female virgin SD rats (n=50) by intravaginal balloon inflation. Then the rats were divid-ed randomly into treatment group (n= 25) and control group (n= 25), treatment group accepted peri-urethral injection of 1.0μg human IGF-Ⅱ to the middle urethral muscle, control group accepted nor-mal saline injection. Five rats in each group were sacrificed at 2, 4, 6, 8, 14 day respectively and the whole urethra specimens were processed for RT-PCR to detect the expression of IGF-Ⅰ ,Ⅱ mRNA. A normal control group (n = 5) was set without intravaginal balloon inflation and injection. Results The expression of IGF-Ⅰ mRNA in control group increased at day 4, 6, 8, 14, the IGF-Ⅰ/β-actin ra-tios were 0. 58±0.15, 1.73±0.31, 2.30±0.29, 0. 46±0. 06. The expression of IGF-Ⅰ mRNA in treatment group increased at all time points, as 0. 69±0.21, 1.45±0.17, 2.25±0.45, 2.90±0.49, 1.92±0. 31. The difference was significant on day 4, 14(P<0.01), and day 8 (P<0.05) compared with the control group. The expression of IGF-Ⅱ mRNA in control group increased at day 4, 6, 8, as 0.42±0. 14, 1.51±0. 59, 1.31±1.04. The expression of IGF-Ⅱ mRNA in treatment group in-creased at day 4, 6, 8, 14, as 1.04±0.23, 1.94±0.29, 1.75±0.41, 0. 81±0.15. The significant difference was noted on day 4 (P<0. 01)compared with the control group. No expression of IGF-Ⅰand Ⅱ mRNA in the normal control group. Conclusions The expression of endogenous IGF-Ⅰ and IGF-Ⅱ mRNA was up-regulated by periurethral injection of IGF-Ⅰ during regeneration period follow-ing urethral sphincter muscle injury in female rat. Our findings suggest that IGF-Ⅰ facilitates the re-generation of the urethral muscles and may play a role in treatment of stress urinary incontinence in-duced by urethral sphincter muscle dysfunction.

Objective To investigate the relationship between Wnt5a gene and E-cadherin or vimentin gene in breast cancer cell line MCF-7. Methods RT-PCR was used to detect the mRNA expression of Wnt5a, E-cadherin and vimentin in breast cancer MCF-7 cells and the normal human mammary epithelial cell line MCF-10A, respectively, and their correlation was analyzed. Results The mRNA expression levels of Wnt5a and E-cadherin in cell line MCF-7 were significantly lower than those in cell line MCF-10A [(16.93± 2.97)%vs. (27.47±2.76) %, (12.97±1.35) % vs. (20.43±2.60) %, both P<0.05]. The mRNA expression level of vimentin in cell line MCF-7 was significantly higher than that in cell line MCF-10A [(16.53±0.85)%(6.33± 2.08) %, P<0.05 ]. In cell line MCF-7, the expression of Wnt5a was positively related to E-cadherin (г=0.997, P<0.05), but it was negatively related to vimentin (г=-0.998, P<0.05). Conclusions The expression of Wnt5a in human breast cancer cell line MCF-7 is significantly lower than that in cell line MCF-10A, which indicates that Wnt5a is a cancer suppressor gene in breast cancer. The expression of Wnt5a in cell line MCF-7 is positively related with E-cadherin, and it is negatively related with vimentin. Wnt5a may cause invasion and metastasis of breast cancer cell through the breast epithelial mesenchymal transitions.