ObjectiveTo establish the urinary proteome profile of the metabolic syndrome ( MetS ) patients,compare the different urinary proteins between the MetS patients and the normal individuals,and analyze the function of the different proteins,so as to explore the pathogenesis of MetS.MethodsOvernight urine were collected from normal controls (n =6) and MetS patients ( n =6).Acetone precipitation method was used to precipitate proteins of urine.Intra-group proteins were mixed together,identified by reversed phase liquid chromatography-mass spectrometry/mass spectrometry and quantified relatively using spectral counting method.The functions of differential proteins were analyzed using Panther.ResultsA total of 807 and 630 proteins were identified respectively in normal controls and MetS patients.Comparing MetS patients with normal controls,sixty different proteins were found,of which 23 proteins were up-regulated and 37 proteins were down-regulated in MetS patients.In the up-regulated proteins,plasminogen was involved in the plasminogen activation cascade and isoform of alphaenolase,phosphoglycerate kinase 1 and fructose-bisphosphate aldolase B down-regulated in MetS patients were involved in the process of glycolysis and fructose metabolism.ConclusionsThe urinary proteome profile of patients with MetS was established by reversed phase liquid chromatography-mass spectrometry/mass spectrometry.Different proteins between MetS patients and normal people were identified.The plasminogen activation cascade,glycolysis and fructose metabolism play key roles in the pathogenesis of MetS.

Objective To investigate the discriminator value of Han Chinese first morning urine albumin creatinine ratio (ACR) for determining the microalbuminuria. Methods A total of 1056 participants (494 males and 562 females) were selected from epidemiologic study of the metabolic syndrome and chronic kidney disease in Pinggu district, Beijing. Eight-hour overnight urinary albumin excretion (UAE) was regarded as the gold standard for defining the albuminuria,and the ROC curve analysis was used to determine the ACR discriminator value for microalbuminuria. Results (1)Microalbuminuria was found in 12.5% of participants,macroalbuminuria in 1.7%. (2)The ACR discriminator value for microalbuminuria by ROC curve analysis was 1.95 g/mol (sensitivity 97.6% and specitivity 88.6%) for men, 3.62 g/mol(sensitivity 83.8% and specitivity 89.1%) for women, 2.78 g/mol (sensitivity 88.7% and specitivity 85.9%)for overall. The upper boundary of microalbuminuria by ROC curve analysis was 22.59 g/mol (sensitivity 100.0% and specitivity 98.8%) . (3)The inter-rater agreement of the result in this study showed that sensitivity was 91.3% and specitivity was 88.2%, positive likelihood ratio was 7.56 and negative likelihood ratio was 0.10, positive predictive value was 56.9% and negative predictive value was 98.4%. Conclusions The ACR discriminator value for determining microalbuminuria is obviously higher in women than that in men, and is higher than recommendation of international guidelines. The result by ROC curve analysis has better sensitivity and specitivity.

Objective To investigate the relationship between the urinary albumin excretion (UAE) and serum uric acid in general population. Methods The study participants were derived from the epidemiological study on the association of metabolic syndrome and chronic kidney disease (CKD) in Pinggu district, Beijing. A total of 992 participants (463 men and 529 women) aged from 30 to 75 years were enrolled in this study. For each participant, UAE, serum uric acid, serum creatinine, and serum lipids were detected and other potential risk factors for CKD were surveyed. Results ( 1 ) The frequencies of microalbuminuria, macroalbuminuria and hyperuricemia were 12.9% , 1.8% and 4.3% respectively. The persons with hyperuricemia had significantly higher frequency of albuminuria than those without hyperuricemia (37. 2% vs 13. 7% , P ＜0. 01). (2) The participants were divided according to the quartiles (25% , 50% , 75% ) of serum uric acid level, and the frequencies of albuminuria in males were 13. 2% , 13. 9% , 17. 2% and 25.4% , while those in females were 8. 4% , 6. 2% , 9. 6% and 24. 8%. ( 3 ) Multivariate logistic regression analysis showed, hyperuricemia was significantly associated with albuminuria in females (OR =2. 31, 95% CI 1. 15-4. 68; P=0.02), but not in males. If the persons with reduced renal function were excluded, similar result still could be gained. Conclusions The prevalence of albuminuria increases gradually with uric acid elevation. Serum uric acid is an independent risk factor of elevated UAE, especially in females.

Objective To elucidate the clinical and pathological characteristics of the patients with thymoma-associated glomerulonephropathy.Methods In this retrospective study,the clinicopathologic characteristics of patients diagnosed as thymoma-associated glomerulonephropathy inPeking University First Hospital during the period between Oct 2008 and Jun 2017 were analyzed,including the histological classfication of thymoma,the clinicopathological features and the short-term prognosis.Results Altogether twelve patients were included with an average age of (55+ 16) years;male/female ratio was 3∶ 1.The B2 type thymoma was the most common type.Nine cases also suffered from myasthenia gravis,and eight cases of glomerulopathy accompanied by thymoma activity.The clinical presentation of glomerulopathy included nephrotic syndrome (11/12),acute kidney injury (10/12).Eleven patients received renal biopsy,among which five cases were minimal change nephropathy,three cases were membranous nephropathy,and the other three cases were focal segmental glomerulosclerosis,thrombotic microangiopathy and endocapillary proliferative glomerulonephritis,respectively.Eleven patients received immunosuppression therapy.After a median 12 months follow up,the proteinuria decreased in 7 cases,and renal function completely or partially recovered in 6 cases.Conclusions Minimal change disease is the most frequent pathological type of thymoma-associated glomerulonephropathy.Immunotherapy with glucocorticoid as first-line drug may be considered for thymoma-associated glomerulonephropathy with surgery,chemoradiation contraindications or non-remission of kidney disease after anti-tumor therapy.

BACKGROUNDChronic kidney disease (CKD) is a common disorder associated with multiple adverse clinical consequences, especially cardiovascular risk and end-stage renal disease. A recent national survey demonstrated that CKD has become a leading health problem in China. There is an urgent need to implement an in-depth investigation of the CKD burden and also to explore underlying mechanisms of CKD progression and it association with adverse consequences.

METHODSThe Chinese Cohort Study of Chronic Kidney Disease (C-STRIDE) is the first national CKD cohort in China. It will enroll approximately 3 000 pre-dialysis CKD patients aged between 18 and 74 years and follow-up for at least 5 years. Questionnaires, anthropometric measures, laboratory tests, and biomaterials will be collected at baseline and annually. The principal clinical outcomes of the C-STRIDE consist of renal disease events, cardiovascular events, and death. Based on the longitudinal clinical data and biomaterials, the risk factors with CKD progression and other outcomes will be analyzed, and candidate markers and predicted models will be established.

CONCLUSIONThe C-STRIDE would provide important evidence for underlying mechanisms of CKD progression, valuable information for clinical guidelines, and healthcare policies in China.

Biomarkers ; China ; Cohort Studies ; Female ; Humans ; Male ; Renal Insufficiency, Chronic

【Objective】 To construct the Rb luciferase reporter gene assay system and detect the activation ability of Rb gene for screening the targeted drugs. 【Methods】 The synthetic Rb gene sequence was annealed to form a double-stranded DNA structure and then inserted into the polyclonal site of pGL6-TA. The junction product was transformed into E.coli DH5α competent cells for expanded culture, and the constructed pGL6-Rb-Luc plasmid and pGL6-TA plasmid were transfected into HEK293 cells. The monoclonal cell line HEK293-Rb-Luc with stable expression was screened by G418, and the activation and inhibition of Rb in HEK293-Rb-Luc were tested by serum and CDK4/6 inhibitor Palbociclib. 【Results】 The sequence of Rb reaction elements in pGL6-Rb-Luc was completely correct. The recovery of serum culture significantly increased the luciferase activity in HEK293-Rb-Luc (P<0.001). Compared with 0 nmol/L, 25, 50, 75 and 100 nmol/L, CDK4/6 inhibitor Palbociclib made the inhibition rate of Rb activity rise to 6.90%, 40.23%, 50.57% and 52.07%, respectively (P<0.05). 【Conclusion】 The Rb luciferase reporter gene detection system HEK293-Rb-Luc was successfully constructed, which can effectively detect the activation level of Rb.

【Objective】 To clone the full-length of human kidney and brain protein (KIBRA) coding sequence in eukaryotic expression vector and provide a model for studying the biological function of KIBRA in breast cancer cells. 【Methods】 Total RNA of human breast cancer cell line MCF7 was extracted. After reverse transcription, the full length of KIBRA (NM_001161661.2) coding region was amplified by PCR, and cloned into eukaryotic expression vector pCMV-Blank. After identification, it was defined officially as pCMV-KIBRA. Then it was transfected into MCF7 cells, and the expression of KIBRA was detected by real-time PCR and Western blotting after 48 hours. The primary, secondary and tertiary structures and post-transcriptional modification sites of KIBRA were analyzed with bioinformatics software. 【Results】 Bacterial PCR, double enzyme digestion and DNA sequencing results showed that the correct sequence of KIBRA was inserted into the vector pCMV-KIBRA. The mRNA and protein expressions of KIBRA were significantly increased in MCF7 cells transfected with pCMV-KIBRA. Bioinformatics analysis showed that KIBRA was composed of 1119 amino acids. There were 52 phosphorylation sites, 1 acetylation site and 5 ubiquitination sites, and the protein structure was mainly α-helix and random coil. 【Conclusion】 The eukaryotic expression vector of full-length of human KIBRA coding sequence was successfully constructed and overexpressed in breast cancer cell line MCF7, which can lay a foundation for studying the biological function of KIBRA in breast cancer.