Objective To evaluate the efficacy of cyclosporine A plus prednisone in the treatment of refractory nephrotic syndrome in children.Methods The 26 refractory nephrotic syndrome children were treated with CsA plus prednisone,3 ～6 mg/(kg d),Po.The duration of treatment was 3 to 27 months (12.69 ± 6.44) mos.The 24 h quantitative urinary proteins,serum cholesterol,urea nitrogen,plasma creatinine,N-acetyl beta amino glycosidase enzymes cystatin C were detected before and after treatment,and the adverse drug reactions were accessed.Results Among 26 cases,12 cases of steroid-resistant NS,6 cases of steroid-dependent NS,and 8 cases of frequent relapse NS were included.16 patients (61.54％) were complete remission,8 patients (30.77％) partial remission,2 cases (7.69％) were non-remission,The total remission rate was 92.31％.The 24 hours urine protein was 3.01 g and 0.63 g before treatment and after treatment,respectively,with a statistically significant difference (P ＜0.01); serum cholesterol was (7.72 ± 3.86) mmol/L and (7.15 ± 3.23) mmol/L; nitrogen urea was (3.93 ± 1.44) mmol/L and (4.04 ± 1.27) mmol/L,creatinine (33.38 ± 13.16) μmol/L and (35.64 ± 3.53) μmol/L serum N-acetyl beta amino glycosidase enzymes was (18.96 ±4.86) u/L and (20.45 ±5.85)u/L before treatment and after treatment,respectively,without a statistically significant difference (P ＞ 0.05).The response to CsA was no significant difference in SRNS,SDNS and FRNS.Children complete remission,follow-up to 6 months,9 months,12 months and 18 months of the recurrence rate was 37.5％,31.25％,18.75％ and 12.5％.Eight cases ended treatment 3 ～ 6 months,all cases were not recurrence.The main adverse effects of CsA included hirsutism,tremble,gastrointestinal reaction and so on,and liver kidney toxicity was not obvious during the therapy course.Conclusions The treatment of CsA in combination with prednisone to children refractory nephrotic syndrome had a significant curative effect,which could obviously minimize the dosage of glucocorticoids and reduce the recurrence after at least one year of maintenance treatment of CsA.

Objective To probe the oprⅠ gene in rat model with Pseudomonas aeruginosa septicemia by FQ-PCR,and compare the sensitivity and specificity between FQ-PCR and traditional germiculture,and check the change of oprI gene before and after the antibiotic therapy as to rapidly judge its sensitivity.Methods The standard Pseudomonas aeruginosa with five different concentration were prepared,the drug-sensitive test wbre used to find lhe sensitive antibiotics.120 SD rats were random divided into five groups,five different concentrations of Pseudomonas aeruginosa were injecked into the rats with the same volume.Six rats of each group were picked up for taking blood for culture at the time points of Oh,12h,24h,and 48h after narcotization.Finally,the oprⅠ gene of each blood samples were checked with FQ- PCR.72 rats were random divided into three groups,therapeutic group,treated group and control group.Pseudomonas aeruginosa with the concentration of 1×109 CFU/ml were injected into those rats.Sensitive antibiotics,insensitive antibiotics and 0.9% NaCl were given to the therapeutic,treated and control group rats respectively.Six rats of each group were picked up for taking blood for culture at the time point of Oh,12h,24h,and 48h after narcotized.Finally,the oprⅠ gent of each blood sample were checked with FQ-PCR.Results The blood culture were positive in each period of the concentrations 1×109 CFU/ml and 1×108 CFU/ml.Results of FQ-PCR showed that the copy number decreased with time going,all of which were positive.The blood culture were positive at the time points of Oh and 12h with the concentrations of 1×107 CFU/ml and 1×106 CFU/ml,were positive with concentration of 107 CFU/ml at the time point of 24h,but negative with concentration of 107 CFU/m at the time point of 48h,and negative with the concentration of 1×106 CFU/ml at the time points of 24h and 48h.The blood culture were negative in each period of the concentration of 1×105 CFU/ml,and the results of FQ-PCR were negative.The blood culture were positive in each period of both treated and control group,but negative in each period of therhpeutic group,all the results of FQ-PCR were positive.Conclusion The coincidence rate between the method of FQ-PCR and trgditional germicuhure were 100%.Though the sensitivity of FQ-PCR was not increased,the time needed by diagnosis was shorter After treated with effective antibiotic,fhe sensitivity of FQ-PER to diagnosis Pseudomonas aeruginosa septicemia was higher than that of traditional germicuhure,and the experiment time was shorter.Detected the changes of the oprⅠ gene copies number may be helpful to estimate the sensitivity of antibiotic.

OBJECTIVE To investigate the effect and mechanism of dihydroartemisinin(DHA)on lipo-polysaccharide(LPS)-induced acute lung injury(ALI)in mice using whole-genome sequencing.METHODS An ALI mouse model was established via intraperitoneal injection of 10 mg·kg-1 lipopolysaccharide.The mice were divided into normal control group(n=10),model group(n=10)and model+DHA group(n=10).The mice in the model+DHA group were injected intraperitoneally with 20 mg·kg-1 DHA,while those in the normal control group and LPS group were injected intraperitoneally with solvent of DHA,saline containing 1%Tween 80 and 10%Macrogol 400.The mice were executed 24 h after drug administration.The wet and dry weight ratio(W/D)of lung tissue was calculated.Hematoxylin-eosin(HE)staining was used to observe histopathological damage in the lung.Classified counts of inflamma-tory cells in alveolar lavage fluid were performed.Enzyme-linked immunosorbent assay(ELISA)was used to detect the levels of interleukin-1β(IL-1β),IL-6,and tumor necrosis factor-α(TNF-α)in alveolar lavage fluid.Real-time quantitative PCR(RT-qPCR)was used to detect mRNA levels of placenta-specific 8(Plac8),Toll-like receptor 7(TLR7),IL-1β,IL-6 and TNF-αin lung tissue.The whole gene transcriptome was sequenced by RNA transcriptome sequencing(RNA-seq)using the Illumina HiSeq high-throughput sequencing platform before the function and signal pathway of differentially expressed gene mRNA between the groups were enriched and analyzed using GO and KEGG enrichment analysis methods.RESULTS Compared with the model group,the lung W/D values of mice,the pathological damage,inflammatory cells in alveolar lavage fluid,expression levels of IL-1β,IL-6 and TNF-α in alveolar lavage fluid(P<0.01,P<0.01,P<0.01),and the mRNA expression levels of IL-1β,IL-6 and TNF-α were significantly reduced in lung tissues in the model+DHA group(P<0.01,P<0.05,P<0.05).Whole gene transcriptome sequencing revealed that immune-related Plac8 and TLR7 genes were significantly upregu-lated(P<0.01)in mouse lung tissue of the model group but significantly downregulated(P<0.05)in mouse lung tissue of the model+DHA group.The results of RT-qPCR of Plac8 and TLR7 verified the results of whole gene transcriptome sequencing.GO and KEGG analysis showed that Plac8 and TLR7 were mainly related to the regulation of cytokine production,T/B cell activation and signal transduction,chemo-kine signal transduction and NF-κB signal transduction.CONCLUSION DHA might reduce LPS-induced lung damage and ameliorate the inflammatory condition in lungs of ALI mice.The mechanism of action may be that DHA negatively regulates the signaling pathways involved in TLR7 and Plac8 by decreasing the expressions of TLR7 and Plac8 mRNA before regulating a series of immune responses such as secretion of inflammation-related cytokines and activation of immune cells,thereby reducing inflam-matory damage in lungs.