AIM: To establish a method of determining the fingerprints of water-soluble composition in Fructus schisandrae chinensis by HPLC. METHODS: Separation was performed on Discovery SB-C18(150 mm?4.6 mm,5 ?m) analytical column with mobile phase consisting of water and acetonitrile with gradient elute at the flow rate of 1.0 mL/min.The UV detection wavelength was set at 203 nm. RESULTS: Compared with reference peak of schisandrin,8 common peaks on frngerprints of Fructus schisandrae chinensis were indicated by the method with satisfied stability,precision and repeatability. CONCLUSION: There are good similarities among 10 batches of Fructus schisandrae chinensis,and this method is reliable,simple and provides a reference standard for the quality control of Fructus schisandrae chinensis.

Objective:To investigate the relationship between VitD 3 concentration and glucose and insulin levels of OGTT in patients with CKD 3-5 stages.Methods: We included the patients with CKD 3 and 4 and 5 stages who fulfill the including standard.All patients were recorded the concentrations of [1,25 (OH):D3]concentration of glucose and insulin at fasting ,postprandial 1 h,2 h during OGTT and concentration of glycosylated hemoglobin level ,C peptide concentration.We performed the correlation analysis about [1,25 (OH):D3],glucose and insulin.Results: We totally included 91 patients with 3-5 stages CKD into our study.The D3 concentration of stage 3 were 160.9-261.3 mmol/L[(218.38±8.67)mmol/L] of stage 3,75.2-166.3 mmol/L[(117.01±4.72) mmol/L] of stage 4 and 11.8-96.5 mmol/L[(41.91±12.83)mmol/L] of stage 5 (P<0.05).The average concentrations of serum glucose at fasting,1 h after the meal and 2 h after the meal was(4.74±0.21)mmol/L,(8.31±0.43)mmol/L and(7.36±0.32)mmol/L in 3 stage and (4.92±0.25) mmol/L,(9.14±0.15) mmol/L and (9.14±0.39)mmol/L at 4 stage and (4.81±0.13)mmol/L, (10.72±0.41)mmol/L and (10.72±0.49)mmol/L at 5 stage (P<0.05).The average concentrations of insulin during OGTT at fasting,1 h after the meal and 2 h after the meal was (6.58±0.32) μU/L,(57.78±5.63)U/L and (42.77±8.45)U/L in 3 stage (6.03±0.53)U/L,(55.69±7.35)U/L and (62.52±5.39)U/L in 4 stage and (6.12±0.65)U/L,(62.82±9.73)U/L and (77.34± 8.62)U/L in 5 stage (P<0.05).Correlation analysis shows that the concentration of 1,25 (OH):D3 of different stages of patients with CKD and vitamin D 3 concentration and glucose tolerance test was found to be inversely associated with the insulin levels ( P<0.05 ) . Conclusion:There are obvious differences of concentration of vitamin D 3 between patients with 3-5 stages of chronic kidney disease (CKD).There also showed a negative correlation relationships between glucose and insulin levels ,and vitamin D3 concentration and glucose and insulin levels at OGTT of patients with 3-5 stages CKD.

Objective To investigate the role of oxidative stress-dependent Rasextracellular signal-regulated kinase (ERK1/2) signaling in aldosterone (ALDO)-induced mesangial cell proliferation. Methods The incorporation of 3H-thymidine (3H-TdR) and cell count were used as the measure of mesangial cell (MC) proliferation.Western blotting was used to detect the activation of Ki-RasA,c-Raf,MEK1/2,ERK1/2 and PI3K. Results Aldosterone significantly induced human mesangial cell proliferation,and anti-oxidant N-Acetylcysteine (NAC),catalase,and super oxide dismutase (SOD) significantly inhibited ALDO-induced mesangial cell proliferation (P＜0.01,respectively).Stimulation by ALDO for 3 h,Ki-RasA,c-Raf,MEK1/2,and ERK1/2 activity increased by 4.05-, 3.62-, 4.52-, and 3.40-fold compared with control group (P ＜0.01,respectively).NAC almost completely blocked ALDO-induced Ki-RasA,c-Raf,MEK1/2,and ERK1/2 activation (P＜0.01,respectively).Ki-RasA siRNA dose-dependently inhibited Ki-RasA expression, ALDO-induced Ki-RasA activation, and mesangial cell proliferation (P ＜0.01,respectively).c-Raf inhibitor GW5074 and MEK1/2 inhibitor PD98059 also reduced ALDO-induced mesangial cell proliferation by 65％ respectvely (P＜0.01).Ki-RasA siRNA had no effect on ALDO-induced PI3K phosphorylation.Combining LY294002 and PD98059 completely blocked ALDO-induced mesangial cell proliferation (P＜0.01). Conclusions ALDO-induced Ki-RasA-c-Raf-MEK-ERK signaling activation is dependent on reactive oxygen species (ROS) production,which mediates ALDO-induced mesangial cell proliferation.Inhibition of both ERK1/2 and PI3K signaling simultaneously completely blocks ALDO-induced mesangial cell proliferation.

Objective To investigate the clinicopathological characteristics and prognosis of Henoch-Schonlein purpura nephritis with diffused endothelial cell proliferation (DEP-HSPN) in children. Methods Data of 8 DEP-HSPN cases in Nanjing Children's Hospital within recent ten years were retrospectively reviewed. The clinicopathological features, efficacy and prognosis were compared between DEP-HSPN cases and 48 cases of non-DEP-HSPN. Non-DEP-HSPN cases were divided into two groups according to the clinical classification or the pathological classification.Results (1) In DEP-HSPN, HSP developed nephritis within 4 to 15 days after the initial onset of purpuric rashes. Hematuria was present in all the 8 patients. The main clinical manifestation of DEP-HSPN was nephritic-nephrotic syndrome (4 cases), nephrotic level proteinuria (3 cases) and acute nephritic syndrome (1 case). Four cases had macrohematuria. Six cases had abdominal symptoms and two cases had arthritis. Pathology of all the cases showed grade Ⅲ-b lesion with diffused endocapillary proliferation and segmental necrotizing lesion of the capillary wall, always accompanied with intraglomerular inflammatory cell infiltration. Crescent was found in 4 cases. (2)Compared to non-DEP-HSPN grades Ⅲ, DEP-HSPN showed a shorter course of disease.Macrohematuria, heavy proteinuria, nephritic-nephrotic syndrome, and segmental necrotizing lesion of capillary wall were more common in DEP-HSPN. Compared to non-DEP-HSPN with nephrotic level proteinuria, DEP-HSPN had a lower rate of crescent. (3) Methylprednisolone pulse therapy in early stage, then prednisone combined with cyclophosphamide were used in the treatment of DEP-HSPN.After an average follow-up period of seven months, one patient showed complete remission, five showed persistent microhematuria, and two showed persistent microhematuria accompanied with minor proteinuria. No significant difference of prognosis was found between DEP-HSPN and nonDEP-HSPN. Conclusions DEP-HSPN has an acute onset. The main clinical manifestation of DEP-HSPN is nephritic-nephrotic syndrome and nephrotic level proteinuria, always accompanied with macrohematuria. Immunosuppressant treatment in the early stage of disease is effective for a short-term outcome.

Objective To investigate the clinicopathological characteristics and treatment of C1q nephropathy in children.Methods Data of 23 C1q nephropathy cases in Nanjing Children's Hospital during recent eight years were retrospectively reviewed. Results The incidence of C1q nephropathy was 4.78％ in primary glomerulonephritis proven by biopsy.Among 23 patients,15 were boys and 8 were girls.The mean age at onset was (5.0±3.4) years old with a range of 0.9-12.4 years.The clinical manifestations included nephrotic syndrome(NS) in 18 cases (78.3％),nephrotic-range proteinuria in 4 cases(17.4％) and microhematuria in 1 case.Two patients with NS and one patient with nephrotic-range proteinuria also presented microhematuria.One patient with NS who received oral herbal medicine for two weeks developed acute renal insufficiency at the same time of diagnosis.Three cases had a family history of kidney disease,among them two patients(presented nephrotic range proteinuria) were siblings,their father had proteinuria as well,and routine genetic examination confirmed familial Denys-Drash syndrome in association with C1q nephropathy.One NS patient's sister had nephrotic-range proteinuria too,but renal biopsy was not performed.No patient had hypertension.None of the patients had low C3 or C4 levels,and serological markers of systemic lupus erythematosus were absent.Light microscopy showedminimalchangedisease (MCD)in13cases (56.5％), mesangialproliferative glomerulonephritis(MsPGN) in 6(26.1％) and focal segmental glomerulosclerosis(FSGS) in 4(17.4％).Immunofluorescence displayed C1q co-deposits of IgG(78.3％),IgM(78.3％),IgA (34.8％) and C3 (47.8％),and a full-house pattern was found in 6 patients (26.1％).Electron microscopy revealed 4 out of 19 had mesangial deposits,except for 4 patients whose glomerulus could not be found.Children with either NS(18 cases) or nephrotic-range proteinuria(2 cases)received prednisone,among them,15 were steroid-resistant,4 were steroid-dependent,only 1 was steroid-sensitive.Those with steroid-resistant(15 cases) or steroid-dependent(3 cases) received further immunosuppression with cyclophosphamide(CTX) or cyclosporine A (CsA).One NS case of steroid-dependent received prednisone re-induction therapy.The siblings associated with DenysDrash syndrome and one case presented microhematuria were commenced on angiotensin-converting enzyme inhibitor(ACEI).Of the 19 patients with sufficient follow-up date,15 cases (78.9％)achieved complete remission,2 cases(10.5％) achieved partial remission,and 2 cases (10.5％) were ineffective.Median follow-up was 15 months.Remission of the NS occurred in 94.4％ (17/18)while nephrotic-range proteinuria was 50.0％(2/4).Remission of MCD was 100.0％,MsPGN was 83.4％(5/6),but FSGS was only 50.0％(2/4).Conclusions C1q nephropathy is rare,and often manifests as steroid-resistant or steroid-dependent NS and nephrotic-range proteinuria.The most common histological feature is MCD,and some as MsPGN or FSGS.A combination of prednisone and immunosuppressive agent is always effective for MCD and MsPGN,but FSGS always has a poor response.

Objective To detect the α1-antitrypsin (AAT) concentration in urine samples of children with primary nephrotic syndrome (PNS) before initiation of glucocorticoid treatment,in order to verify whether it could predict the response to glucocorticoid-based therapy.Methods Forty-three children diagnosed as PNS initially were chosen as subjects,namely steroid-sensitive nephrotic syndrome (SSNS) and steroid-resistant nephrotic syndrome (SRNS) depending on reaction to glucocorticoid therapy four weeks later,and 15 healthy children serving as normal control.The mid stream of the first morning urine samples were collected from children before taking glucocorticoid.ELISA kit was used to quantify the urinary AAT concentration which was revised by urine creatinine further.The data of urine AAT/Cr were expressed as median with interquartile range.Data analysis was performed using the SPSS 17.0.Results AAT was absent in urine samples of normal healthy children,and there were no statistic differences of the AAT concentrations in urine between children with SSNS and SRNS [(30.4+4.5) mg/L vs (31.8+4.6) mg/L,t=-1.0,P=0.33].The level of urine AAT/Cr in children with SRNS was higher than that in children with SSNS [0.049(0.028-0.073) vs 0.028(0.022-0.036),Z=2.4,P=0.02].Among the laboratory parameters of the two subgroups before taking glucocortiod,the levels of platelet,blood white cell count,serum globulin,urine white cell count,urine red cell count,urine IgG and urine α1-microglobulin were significantly different (P＜0.05).Three parameters that included urine AAT/Cr (OR=6.81 × 1028,P=0.O05),serum globulin (OR=1.69,P=0.01) and urine α1-microglobulin (OR=1.05,P=0.009) further entered the logistic regression model to predict the SRNS independently.The ROC curve based on the level of the urine AAT/Cr was constructed,and the area under the curve (AUC) was 0.72.When the cutoff value of urine AAT/Cr was 0.035,the sensitivity and specificity of the urine AAT/Cr prediction were 68％ and 75％ respectively (Youden' s index 0.43).The AUC that based on the logistic regression model which included urine AAT/Cr,serum globulin and urine α1-mieroglobulin was improved to 0.94,and the sensitivity and specificity of the model prediction were 95％ and 83％ respectively (Youden' s index 0.78).There was no significant difference of the urine AAT/Cr level among the different pathological types of the children undergoing renal biopsy.Conclusions There are no statistic differences of the AAT concentrations in urine between children with SSNS and SRNS.The level of urine AAT/Cr is significantly higher in the SRNS than that in the SSNS which can be as a candidate biomarker to predict the response to glucocorticoid-based therapy.It has a better prediction efficacy based on the model which includes urine AAT/Cr,serum globulin and urine α1-microglobulin.

Objective To investigate the relationship of red cell distribution width (RDW) with all-cause mortality and cardiovascular disease (CVD) mortality in patients undergoing maintenance hemodialysis (MHD).Methods A retrospective analysis was performed in patients who initiated MHD from January 2008 to September 2017 in the hemodialysis center of the Second Affiliated Hospital of Soochow University.Basic data on demographic,dialysis and laboratory were collected,and echocardiography indicators and clinical outcomes were recorded.Patients were divided into four groups according to the quartile of RDW level.Kaplan-Meier survival analysis was used to compare the difference of survival rate among the groups.Cox regression analysis was used to analyze the risk factors of all-cause and CVD-related mortality,and predictive value of RDW for all-cause and CVD-related death in hemodialysis patients.Results A total of 268 MHD patients were enrolled in this study with age of (60.9± 15.8) years and dialysis duration of (58.1±9.1) months,including 159 males (59.3％).Kaplan-Meier survival analysis showed that the 1-year overall survival rates of Q1 group (RDW≤ 13.8％,n=61),Q2 group (RDW 13.9％-14.6％,n=66),Q3 group (RDW 14.7％-15.6％,n=73)and Q4 group (RDW≥15.7％,n=68) were 96.8％,95.1％,93.1％ and 85.7％ respectively;3-year overall survival rates were 88.5％,87.5％,59.2％ and 51.8％ respectively;5-year overall survival rates were 71.5％,65.4％,33.6％ and 17.7％ respectively;The difference between the groups was statistically significant (all P ＜ 0.01).The 1-year CVD survival rates were 98.4％,96.6％,95.8％ and 92.4％ respectively;3-year CVD survival rates were 94.8％,92.5％,84.4％ and 70.4％ respectively;5-year CVD survival rates were 86.9％,81.3％,65.6％ and 51.3％ respectively;The difference between the groups was statistically significant (all P ＜ 0.01).Multivariate Cox regression analysis showed that RDW≥15.7％ was an independent risk factor for all-cause and CVD-related mortality in MHD patients.The risk of all-cause mortality in Q4 group was 3.098 times higher than that in Q 1 group (95％ CI 1.072-8.950,P=0.037) and the risk of CVD-related mortality was 2.661 times (95％ CI 1.111-8.342,P=0.048).Receiver operating characteristic curve (ROC) showed that RDW=14.85％ was the best cut-off point for predicting the all-cause mortality in HD patients (P ＜ 0.01),RDW=15.45％was the best cut-off point for predicting the cardiovascular disease mortality (P ＜ 0.01),and RDW=14.45％ had a higher 5-year survival rate (P ＜ 0.01).Conclusion RDW can independently predict all-cause and CVD-related mortality risk in hemodialysis patients,and it has important value for prognosis.

To investigate the impact of goal directed analgesia on the outcome of patients with mechanical ventilation in intensive care unit. A total of 126 patients who needed mechanical ventilation were recruited. With a method of before and after paired comparison, they were divided into two group:( 1 ) analgesia with empirical administration or control group; ( 2 ) goal directed analgesia based on critical-care pain observation tool (CPOT). Compared with the control group, after goal directed analgesia was applied, the consumption of midazolam significantly dropped from ( 368. 47 ± 27. 41 ) mg to ( 151. 27 ± 29. 31 ) mg (P<0. 05), whereas the consumption of dexmedetomidine significantly increased from ( 623. 62 ± 20. 91) μg to (812. 34 ± 22. 57) μg(P<0. 05). The median score of Richmond agitation-sedation scale increased from -3 to -1. The incidence of delirium significantly reduced from 23. 81% to 17. 46%( P<0. 05). The mean ventilator duration was significantly shortened from (168. 49 ± 11. 41) h to (142. 38 ± 13. 24) h(P<0. 05). ICU length of stay was significantly shortened from (23. 64 ± 9. 26) d to (19. 63 ± 8. 46) d ( P < 0. 05 ) . Due to the mild sedation, patients receiving goal directed analgesia report less delirium, less ventilation time and shorter ICU length of stay, suggesting that the general outcome is improved.

Objective: To explore the differential expression profiles of DNA methylation sites/regions and potential molecular mechanisms in the peripheral blood of coronary heart disease (CHD)-induced unstable angina pectoris patients with or without Qi deficiency and blood stasis syndrome, and to provide scientific evidence for the conbination of disease and syndrome. Methods: According to the pre-determined inclusion and exclusion criteria, the study subjects were enrolled and divided into two groups namely CHD-induced unstable angina group (G group) and healthy control group (J group) to conduct “disease” analysis, while G group was further divided into Qi deficiency and blood stasis syndrome group (case group) and non-Qi deficiency blood stasis syndrome group (control group) to perform “syndrome” analysis. The general data and clinical information of the study subjects were collected. The peripheral venous blood was extracted on an empty stomach, and the Illumina Infinium MethylationEPIC BeadChip (850K methylation chip) was used to detect the differential expressionprofiles of DNA methylation in each group, ChAMP software (V 2.14.0) was used for the differential methylation data analysis, with a threshold of the adjusted P value (adj.P.val) < 0.01. Gene Ontology (GO) and Kyoto Encyclopedia of Genomes (KEGG) were employed for the functional and pathway enrichment analyses of related mapped genes. Results: A total of 263 differentially methylated CpG positions (DMPs) were screened out between G and J groups, including 191 hypermethylated positions such as cg05845204 and cg08906898, and 72 hypomethylated positions such as cg26919182 and cg13149459. These positions were mainly mapped to 148 genes encompassing RNA binding motif protein 39 (RBM39), acetyl-CoA acyltransferase 2 (ACAA2), protein phosphatase 1 regulatory subunit 12B (PPP1R12B), and the dual-specificity tyrosine phosphorylation-regulated kinase 2 (DYRK2). GO functional enrichment analysis revealed that the genes of the DMPs were primarily enriched in protein localization to chromosomes, regulation of cell morphogenesis, negative regulation of calcium-mediated signals, etc. KEGG pathway analysis suggested that the genes were mainly enriched in fatty acid metabolism and endocytosis pathways. In addition, a total of 23 differential methylation regions (DMRs) were identified, with overlapping genes such as transmembrane protein 232 (TMEM232), ribosomal protein large P1 (RPLP1), peroxisomal biogenesis factor 10 (PEX10), and forkhead box N3 (FOXN3) recognized. It was found that GO functions were mainly enriched in the negative regulation of Ras protein signal transduction, small GTPase-mediated signal transduction, negative regulation, etc. A total of 1 703 differential methylation sites were screened out between case and control groups, including 444 increased methylation positions such as cg05573767 and 1 259 decreased methylationpositions such as cg19938535, and cg03893872. These positions were mapped to 1 108 genes such as ribosomal protein S6 kinase A2 (RPS6KA2), leucine rich repeat containing 16A (LRRC16A), and hedgehog acyltransferase (HHAT). According to the GO functional enrichment analysis, the genes relating to the DMPs were mainly enriched in biological functions such as transmembrane receptor protein serine/threonine kinase signaling pathway and axonogenesis. The KEGG pathway enrichment analysis suggested the involvement of Rap1 signaling pathway, adenosine 5’-monophosphate-activated protein kinase (AMPK) signaling pathway, etc. A total of 21 DMRs were identified, including 22 overlapping genes such as mucin 4 (MUC4), three prime repair exonuclease 1 (TREX1), and LIM homeobox 6 (LHX6). GO analysis demonstrated that the genes primarily participated in molecular functions such as positive regulation of transmembrane transport, regulation of fatty acid metabolism, and copper ion binding. Conclusion This study reveals the methylation patterns of DMPs and DMRs in patients with Qi deficiency and blood stasis syndrome caused by CHD-induced unstable angina pectoris. Potential epigenetic regulation of fatty acid metabolism, Rap1 signaling, and other molecular functions are involved in the development of CHD between the "disease" and "syndrome".