Objective    To explore the level of 25-OH vitamin D3  in patients with lupus nephritis (LN) and its association with the activity of disease.    Method    Clinical data of 154 patients with lupus nephritis in our hospital from January 2008 to May 2015 were collected. Another 63 healthy adults were recruited as normal controls.    Results    The level of 25-OH vitamin D3 in patients with LN was significantly lower than that in normal controls (P﹤0.01). The concentration of serum 25-OH vitamin D3 was negatively correlated with 24 h urinary protein (r=-0.18, P=0.02), antinuclear antibody (r=-0.20, P=0.01), SLEDAI scores (r=-0.20, P﹤0.01), while positively correlated with serum complement C3 (r=0.32, P﹤0.01). The'full-housed' patients with all 5 antibodies positive in renal biopsy had lower level of 25-OH vitamin D (P＜0.05).    Conclusion    The level of 25-OH vitamin D3 in patients with LN is inversely correlated with the disease activity.

Objective    To analyze the influence factors of serum high-sensitivity cardiac troponin T (hs-cTnT) for non-dialysis chronic kidney disease (CKD) patients, and to further investigate cardiac and renal effects on hs-cTnT.    Methods    Cross-sectional study was applied. Clinical data of 577 non-dialysis CKD patients were collected. Comparison between groups and lineal regression analysis were utilized to investigate the influence factors of hs-cTnT.    Results    Median level of hs-cTnT was 0.013 (0.007-0.029) μg/L, with 1.7% undetectable (hs-cTnT＜0.003 μg/L), and 46.4% greater than 99th percentile (hs-cTnT﹥0.014 μg/L) of the general population. Multivariate linear analysis displayed that higher Ln hs-cTnT was significantly associated with older age, male, diabetes, higher Cys C, higher urine albumin-to-creatinine ratio (UACR), lower estimated glomerular filtration rate (eGFR) and higher LVMI (P＜0.05).     Conclusions    Traditional and non-traditional risk factors of CKD-cardiovascular disease are shown to be associated with serum hs-cTnT level. Cardiac and renal injury may be associated with elevated hs-cTnT among non-dialysis CKD patients.

Objective    To investigate the clinical and pathological features of idiopathic membranous nephropathy (IMN) in young patients.    Methods    Clinical data of 20 young patients, 16 to 44 years, who were diagnosed as IMN admitted to the Zhejiang Provincial People's Hospital from January 2013 to December 2014 were retrospectively analyzed, comparing to 55 mid-aged patients who were diagnosed as IMN during the same period in the hospital. Clinical and pathological features of above mentioned patients were analyzed.    Results    Young patients with IMN accounted for 26.7% of IMN patients. Compared to mid-aged patients, young patients with IMN had lower proportion of hypertension (P=0.003), lower blood glucose level (P=0.010), higher estimated glomerular filtration rate (eGFR) and low-density lipoprotein (LDL) (P=0.012, P=0.038), and lower levels of T3 and T4 (P=0.030, P=0.034). Furthermore, there were less sclerosis glomeruli (P＜0.001), hyaline change of arteriole (P=0.040) and arteriolar wall thickening (P＜0.0001), lower positive ratios of IgA (P=0.008), and more without renal tubulointerstitial lesions (P=0.018) in young patients. There were no statistically significant differences between these two groups in other index.    Conclusions    Compared to mid-aged patients, young patients with IMN have better blood pressure and blood glucose level, higher glomerular filtration rate and LDL. Moreover, thyroid function is significantly affected, meanwhile the lesions of glomerular, interstitial and vascular are mild in young patients.

Objective    To evaluate clinical application value of polymerase chain reaction (PCR) detection for bacteria in peritoneal dialysis associated peritonitis (PDAP).    Methods    Peritoneal dialysis fluid specimens were collected from January 2014 to December 2014 in The First Affiliated Hospital of Anhui Medical University. Conventional bacterial culture and PCR detection were used respectively. According to the bacterial 16S rRNA gene, universal primers were devised and designed, based on reference, the specific primers of 17 kinds of experimental bacteria. Real-time fluorescent PCR (Real-time PCR, qPCR) amplification was implemented. The establishment of standard strain DNA extract was used as positive control; sterile double distilled water was used as negative control. Results    (1) The traditional bacterial culture results showed that positive proportion was 26/40 in specimen of 40 cases, gram-positive strains accounting for 18/26. Main species were epidermis staphylococcus (5/26), hemolysis staphylococcus (4/26), escherichia coli (4/26), and streptococcus viridans (3/26). (2) The PCR detection results showed that total positive rate was 33/40 in 40 patients specimens, among which 2 cases of positive samples ended up with no specific strains being detected; the main bacteria strains in PCR were not different from ordinary culture results. (3) With bacterial culture as the gold standard, the detection sensitivity of PCR technology for PDAP pathogenic bacteria was 96.15% and specificity was 42.86%; the detection positive rate was significantly higher than ordinary culture method. (4) PCR technology for detecting pathogenic bacteria could produce results within 4-6 hours, while reported positive results in the traditional bacterial culture would take (77.88±15.53) hours, which was significantly longer than PCR.    Conclusion    Compared with traditional bacteria culture method, PCR method is more sensitive, simple, and quick. Bacteria detection using PCR technique is of clinic applied value in PDRP.

Objective    To investigate the changes of thyroid function and carotid atherosclerosis in patients on maintenance hemodialysis (MHD).    Methods    A total of 110 stable MHD patients undergoing hemodialysis for at least three months were enrolled in the study. Serum free-T3 (FT3), free-T4 (FT4) and thyroid stimulating hormone (TSH) concentrations were measured by electrochemiluminescence．Plasma levels of homocysteine (Hcy) and C-reactive protein (CRP) were detected. Clinical data and biochemical indicators were collected. These patients were divided into thyroid dysfunction group and euthyroidism group. Prevalence of atherosclerosis was detected by carotid ultrasonography. The associations between the changes of thyroid function and carotid atherosclerosis were analyzed by Logistic regression model.    Results    Among these 110 patients, 42 (38.18%) patients had thyroid dysfunction. Hcy and CRP concentrations were significantly higher in thyroid dysfunction group than those in euthyroidism group (P＜0.05). The intima-media thickness, number of plague and arteriostenosis of carotid were higher in thyroid dysfunction group than those in euthyroidism group (P＜0.05). Multivariate logistic regression analysis showed that increased Hcy and CRP, decreased serum FT3 were independent risk factors for carotid atherosclerosis.    Conclusions    Thyroid dysfunction with low serum FT3 is frequently found in MHD patients. In MHD patients, FT3 is closely correlated to carotid atherosclerosis.

Objective    To explore the effect of hypoxia on exosomes secreted by renal tubular epithelial cells and the function of exosomes in chronic kidney diseases.    Methods    (1) The supernatant of renal tubular epithelial cells which were cultured in normoxia (21% O₂) or hypoxia(1% O₂) for 48 h was collected and centrifuged gradiently to harvest exosomes. Exosomes were identified and compared by transmission electron microscope, nanoparticle tracking analysis, Western blotting and measurement of the protein concentration. (2) Primary peritoneal macrophages of rats were co-cultured with exosomes in different concentrations (1, 10, 50, 100, 300 mg/L). The expression of interleukin-6(IL-6), tumor necrosis factor-α (TNF-α), inducible nitric oxide synthase (iNOS) in cells and supernatant were separately detected by quantitative real-time PCR (qRT-PCR) and ELISA, and the expression of phospho (p)-STAT/STAT and suppressors of cytokine signaling  1 (SOCS1) in macrophages was detected by Western blotting. At last, the expression of inflammatory microRNAs(miR) in exosomes was measured by qRT-PCR.    Results    (1) The vesicles harvested by gradient centrifugation were less than 150 nm and expressed CD63 which was characteristic of exosomes. Hypoxia had no effect on the morphology of exosomes, but stimulated their secretion. (2) Hypoxic exosomes dose-dependently improved the expression of IL-6, TNF-α, iNOS in macrophages polarized by lipopolysaccharide (LPS)  and increased the expression of p-STAT while decreased the expression of SOCS1 (P＜0.01). MicroRNAs referred to inflammation such as miR-155 and miR-27a increased in hypoxic exosomes compared to that in normoxic exosomes (P＜0.05).    Conclusions    Hypoxia makes exosomes promoted the polarization of macrophages to M1, which may account for the microinflammation in chronic kidney diseases.

Objective    To evaluate the effect of telmisartan on serum adiponectin and urine microalbumin(mAlb) level by administrating it to mice with simple obesity, so as to explore new therapies for obesity-related kidney diseases.    Method    A total of 24 8-week-old male OB mice and 8 8-week-old male C57 mice were selected for this study. The genetic background of OB mouse was C57 mouse, but the lepin gene was deleted in OB mouse. OB mice were randomly divided into 3 groups by body weight and fed with high-fat diet for 12 weeks: model group (M group), telmisartan group (T1 group) and losartan group (T2 group). C57 mice acted as control and were fed with general diet for 12 weeks. Serum adiponectin and blood glucose levels were measured before and after treatment. 24 h urine was collected to measure urine mAlb. Quantitative real-time PCR (qRT-PCR) was used to measure the expression level of peroxisome proliferators-activated receptors gamma mRNA(PPARγ mRNA). HE stain was used to observe the morphological changes of kidney and measure the glomerular diameter. Body weight, serum adiponectin level, blood glucose level, urine mAlb level, expression level of kidney PPARγ mRNA, kidney wet weight and glomerular diameter of 4 groups were compared and a correlation analysis was carried out by Person correlation coefficient.    Results    Compared with C group, the urine mAlb level in M group increased (P＜0.01), serum adiponectin level and the expression level of kidney PPARγ mRNA in M group decreased (P＜0.01); The urine mAlb level was negatively correlated with serum adiponectin level and expression of kidney PPARγ mRNA (r=-0.773,P＜0.01; r=-0.469, P＜0.01). The urine mAlb level in T1 group was lower than M group (P＜0.05), serum adiponectin level and expression of kidney PPARγ mRNA in T1 group were higher than M group(P＜0.05). Compared with T2 group, the urine mAlb level in T1 group decreased, serum adiponectin level and expression of kidney PPARγ mRNA in T1 group increased (P＜0.05); Compared with M group, the urine mAlb level in T2 group decreased (P=0.01). The morphological changes of kidney: glomerular volume increase and focal segmental sclerosis were found in some mice in M group and T2 group. No glomerular volume increase and focal segmental sclerosis were observed in T1 group. Conclusions    Telmisartan can reduce urine microalbumin, whose mechanism might be that telmisartan can active the PPARγ and promote the level of serum adiponectin.

Objective    To explore the effect and mechanism of acidification on calcification in chronic renal failure rats.    Methods    In vivo, male SD rats (n=22) were randomly divided into sham operated group (n=6), 5/6 nephrectomy (Nx, n=5), 5/6 Nx+calcitriol group (n=5), 5/6 Nx+calcitriol +acidosis group(n=6). Vascular calcification was determined by von Kossa stain and quantification of calcium. L-type calcium channels(LTCC)β₃ subunit and runt-related transcription factor 2 (runx2) in aortic were measured by immunohistochemistry. In vitro, vascular smooth muscle cells (VSMCs) were primarily cultured and calcification was induced by β-glycerophosphate(β-GP). They were then randomly divided into control +pH7.4 group, high phosphorus +pH7.4 group (10 mmol/L β-GP), high phosphorus +pH7.1 group (10 mmol/L β-GP) and verapamil intervention group (10 mmol/L β-GP+20 mmol/L verapamil). Calcium deposition was measured by Alizarin red staining and quantification of calcium; and alkaline phosphatase (ALP) activity was measured by enzyme linked immunosorbent assay. RT-PCR and Western blotting were used to observe the expression of VSMCs LTCCβ₃ subunit and runx2 mRNA and protein respectively.    Results    In vivo, compared to that in 5/6 Nx+calcitriol group, calcification was significantly reduced in 5/6Nx+calcitriol +acidosis group (P＜0.05); the expressions of LTCCβ₃ subunit and runx2 were obviously down-regulated by acidification (P＜0.05). In vitro, compared to that in high phosphorus +pH7.4 group, calcification was significantly reduced in high phosphorus+pH7.1 group (P＜0.05), as well as ALP activity (P＜0.05); the expressions of runx2 and LTCCβ₃ subunit were down-regulated in high phosphorus +pH7.1 group (P＜0.05). Calcium influx in VSMCs was partially blocked during acidification (P＜0.05). The calcification level and expression of runx2 in verapamil intervention group were lower than that in high  phosphorus +pH7.4 group (P＜0.05).    Conclusion    One of the possible mechanisms of the inhibitory effect of acidification on VSMCs calcification is that acidification prevents LTCCβ₃ subunit expression and calcium influx, leading to the degradation of VSMCs phenotype transforming.

Objective    To investigate a possible molecular mechanism of MiRNA-130b-3p involved in renal damage.    Methods    Human renal tubular epithelial cells (HK-2) were transfected with MiR-130b-3p mimics or normal control mimics. Then HK-2 cells were stimulated with 10 μg/L recombinant TGF-β1 for 72 h. After 72 h, the mRNA and protein expression of Collegen Ⅳ, α-smooth muscle actin (α-SMA), Collegen Ⅰand E-cadherin were quantified by real-time PCR and Western blotting. The mRNA and protein expression of ERBB2IP and PPARγ were also detected. The reporter plasmids containing ERBB2IP 3'-UTR and PPARγ 3'-UTR were constructed. The activity of ERBB2IP and PPARγ were detected by dual luciferase report system.    Results    Compared to NC mimic group，transfection of HK-2 cells with MiR-130b-3p mimics resulted in significantly increased expression of mRNA and protein of Collegen Ⅳ, α-SMA, Collegen Ⅰ, and decreased expression of E-cadherin after stimulating by TGF-β1 (all P＜0.05). And MiR-130b-3p mimic could significantly down-regulate the mRNA and protein expression of ERBB2IP and PPARγ in HK-2 cells (all P＜0.05) whether in the presence of TGF-β1 or not. The dual luciferase reporter assay showed that MiR-130b-3p induced decreased ERBB2IP 3'-UTR luciferase activity compared to NC mimic group, but there was no significant difference between NC mimic group and mut-MiR-130b-3p mimic group. MiR-130b-3p mimic+mut-PPARγ-3'UTR cotransfection group had lower PPARγ luciferase activity than NC mimic + mut-PPARγ-3'UTR group , and MiR-130b-3p+PPARγ-3'UTR group got lower further (all P＜0.01). Conclusions    MiR-130b-3p promotes epithelial-to-mesenchymal transition (EMT) in renal tubular epithelial cells by directly targeting at the 3'-UTR of ERBB2IP and PPARγ, which may play an important role in renal damage of early stage lupus nephritis.