AIM: To test the hypothesis that locally produced renal endothelin(ET) and nitric oxide(NO) play more important role in initiating proteinuria than circulating ET and NO. METHODS: (1) Rat nephrotic syndrome model was made by single injection of adriamycin, and the urinary protein excretion per 24 hours for every two days was measured. (2) Rat plasma and renal cortex were collected respectively 4 days, 8 days, 32 days and 56 days after the injection, and renal cortex homogenate was further prepared; ET of both plasma and renal cortex homogenate were determined by radioimmunoassay; NO of both plasma and renal homogenate were measured by the method of Griess. (3) The correlation between urinary protein and ET or NO concentration was analyzed respectively. RESULTS: (1) The urinary protein excretion of normal rat was 4-7 mg/d, that in nephrotic rat at the 4 time points was 5, 10, 241 and 201 mg/d, there was a significant increase of that at day 8, peaked at day 32, and less decrease at day 56. (2) The plasma ET in nephrotic rats of the 4 time points was 134, 150, 538 and 445 ?g/L, respectively, which of day 8, day 32 and day 56 shown significant higher than that of normal control (126-129 ?g/L). (3) The ET in renal cortex homogenate of nephrotic rat of the 4 time points was 364 6, 652 3, 1 526 8, 1 393 6 pg/g, all of that was much more higher than that of normal rat [(235 9-246 1) pg/g,P

Objective To construct the miRNA‐21 eukaryotic overexpression vector pmR‐21 and to explore its regulation effect on the expression of c‐myc gene in HepG2 .2 .15 cells .Methods The miRNA‐21 precursor gene fragment pre‐miRNA‐21 was amplified by PCR ,then connected to the pmR‐mCherry plasmid vector after double enzyme digestion ,the accuracy of the recombi‐nant vector was verified by double enzyme digestion and sequencing ;then the recombinant vector was transfected into HepG2 .2 .15 cells ,the fluorescent protein expression was observed under the fluorescence microscopy at 24 h and the transfection efficiency was detected by flow cytometry ;the expression of miRNA‐21 was evaluated by real‐time quantitative PCR;at 72 h after transfection ,the expression levels of c‐myc gene were detected by RT‐PCR and Western blot ;CCK‐8 was used to detect the cell proliferation in each group .Results The double enzyme digestion and Western blot verified that the target gene fragment was inserted into the pmR‐mCherry vector;at 24 h after transfection ,intracellular strong fluorescence was seen ,the transfection efficiency was higher than 50% ;miRNA‐21 expression level of the pmR‐21 recombinant vector group was significantly increased;c‐myc gene expression was increased in the pmR‐21 recombinant vector group at 72 h after transfection ,the cell proliferation in the pmR‐21 recombinant group was faster than that in the control group(P<0 .05) .Conclusion The pmR‐21 eukaryotic overexpression vector is successfully con‐structed ,this recombinant vector can express miRNA‐21 stably ;miRNA‐21 can up‐regulate c‐myc gene expression ,c‐myc gene is one of miR‐21′s targets for playing a cancer‐promoting action .

Objective To compare different diagnosis values of age-specific procalcitionin (PCT) and C-reactive protein (CRP) on early-onset neonatal sepsis (EONS) during the first 72h of life.Methods From September 2008 to December 2015,96 neonates (including 2 confirmed sepsis and 94 clinical sepsis) without severe complications were chosen as the EONS group and 170 non-infectious newborns as the control group.A total of 605 blood samples were collected from all 266 newborns.Serum concentration of PCT and CRP were measured in both the EONS group and the control group at each age over the first 72 h of life.The diagnostic value of PCT and CRP within 1 ～ 12 h and 13 ～ 24 h and 25 ～ 48 h and 49 ～ 72 h of life was evaluated by calculating the cut-off values,sensitivity,specificity,and the area under the receiver operating characteristic curve (ROC).Results PCT and CRP levels of neonates within each age in the EONS group were significantly higher than those in the control group during the first 72h of life.(all P ＜ 0.05).Within 1 ～ 12 h,13 ～ 24 h,25 ～ 48 h and 49 ～ 72 h of life,the cutoff value of PCT was 0.45 μg/L (sensitivity 84.2％,specificity 74.4％),1.885 μg/L (sensitivity 73.5％,specificity 75％),0.995 μg/L (sensitivity 82.4％,specificity 74.1％) and 0.51 μg/L (sensitivity 83.3％,specificity 79.2％) respectively;that of CRP3 185 mg/L (sensitivity 68.4％,specificity 82.1％),6.29 mg/L (sensitivity 58.8％,specificity 89.7％),8.615 mg/L (sensitivity 54.3％,specificity 93.9％) and 10.27 mg/L (sensitivity 59.1％,specificity 100％) respectively,and the area under ROC of PCT for the diagnosis of EONS was 0.767,0.754 and 0.755,and 0.8 respectively;that of CRP 0.773,0.8,0.815 and 0.789 respectively.Conclusions There are age-specific cut-off values of PCT and CRP in the diagnosis of EONS without severe complications during the first 72 h of life.Both PCT and CRP are moderately accurate for the diagnosis of EONS.PCT may be more helpful for the early diagnosis of EONS for its higher sensitivity but CRP presents higher specificity.