Objective:To analyze the clinical characteristics of pediatric atypical hemolytic uremic syndrome (aHUS), and provide clinical experience for the diagnosis and treatment of aHUS in China.Methods:It was a single-center retrospective study. Fifteen aHUS children treated and having complete clinical data at Children's Hospital of Nanjing Medical University between December 31, 2017 and October 15, 2023 were enrolled to analyze the clinical features covering laboratory examinations, genetic testing results, and clinical manifestations. The children were classified based on genetic testing and complement factor H (CFH) antibody detection results to analyze the corresponding clinical characteristics.Results:Among the 15 aHUS patients. There were 8 males and 7 females. The onset age was 5.1 (0.7, 10.8) years old. All patients underwent genetic testing, with 9/15 of aHUS-related gene mutation, revealing 2 de novo mutations in complement factors-related genes. Among 11 patients screened for CFH antibody, 6 tested positive. C3 was detected in 14 patients , and C3 decreased in 9 patients. In laboratory examinations, there were notable decreases in red blood cell (RBC) count in 13 patients, platelet (PLT) count in 15 patients, hemoglobin (Hb) in 15 patients and estimated glomerular filtration rate (eGFR) in 14 patients. Blood urea nitrogen (BUN) and serum creatinine (Scr) were markedly elevated in 13 patients and 9 patients, respectively. Twelve patients exhibited elevated transaminase levels, and 14 patients exhibited elevated lactate dehydrogenase (LDH) levels. Clinically, 11 patients had triggers, and 4 patients had clear family histories. Common clinical features including anemia, thrombocytopenia, proteinuria and hematuria were in 15 patients. There were statistically significant differences in RBC count ( Z=-2.84, P=0.005), PLT count ( Z=-6.65, P<0.001), Hb ( t=-3.71, P=0.002), LDH ( Z=3.76, P=0.002), BUN ( Z=2.71, P=0.017), and eGFR ( Z=-3.65, P=0.003) before and after treatment except alanine transaminase, aspartate transaminase, Scr and complement C3 (all P>0.05). There were no significant differences in onset age, RBC count, PLT count, Hb, LDH, alanine transaminase, aspartate transaminase, Scr, BUN, eGFR, and C3 between aHUS-related gene mutation and non-mutation groups, and CFH antibody-positive and negative groups (all P>0.05). Conclusions:aHUS is marked by severity, and has diverse clinical manifestations. There are no significant differences in clinical presentation at admission between hereditary and acquired aHUS, highlighting the critical importance of genetic testing and complement-related factor detection in diagnosing aHUS etiology. The family history plays a supportive role in diagnosis of aHUS.

Objective The aims of this study were to investigate the expression of DKK2,a WNT signaling pathway regula-tor,in nephroblastoma cells and tissues of children,the effect on the proliferation of nephroblastoma SK-NEP-1 cells,and to explore its mechanism. Methods The relative expression of DKK2 in nephroblastoma cells and tissues was analyzed by qRT-PCR and im-munoblotting assays. Overexpressing DKK2 SK-NEP-1 cells were set as the experimental group( DKK2 group);the blank control plasmid group was set as a control group( Vector group),transfected with pcDNA3. 1 ( +) -Flag-DKK2 plasmid( Experimental group)and pcDNA3. 1( +) -Flag-Vector plasmid(Control group). The over-expression of DKK2 was confirmed in SK-NEP-1 cells by RT-PCR and immunoblotting. CCK-8 and cell cloning assays were used to determine the effect of DKK2 on cell prolifera-tion;flow cell cycle and apoptosis assays were used to confirm the effect on cell proliferation in overexpressed DKK2 cells. The xen-graft formation assay in nude mice was to verify the effect of DKK2 on proliferation in overexpressed DKK2 cells;the mechanism of DKK2 in inhibitory proliferation was analyzed by qRT-PCR,Western blotting and immunohistochemistry. Results Compared with normal renal epithelial tissues,DKK2 mRNA was down-regulated in children with nephroblastoma,and the difference was statistically significant(P<0. 001). Compared with the control group,transfected DKK2 cell viability was significantly inhibited after treatment for 24,48 and 72 h( P<0. 05),cell clone formation in the experimental group was significantly inhibited(31. 11% ± 2. 14% ) ( P<0. 05),the cell cycle in the experimental group was significantly arrested at the G1 phase(P<0. 001),and the apoptosis rate in the experimental group was significantly increased(P<0. 001). Compared with the control group,the tumor weight and volume in nude mice were significantly low in the experimental mice which were injected DKK2 overexpression cells(P<0. 001). Active-β-cate-nin and downstream genes were significantly inhibited in over-expressed DKK2 SK-NEP-1 cells. Conclusion DKK2 is down-regulated in human cutaneous nephroblastoma and participates in the mechanism of nephroblastoma by antagonizing Wnt/β-catenin signaling pathway.

Objective To optimize the exposure parameters and reduce the irradiation dose level in infants and young children during digital radiography (DR) chest radiography under the premise of satisfied image quality.Methods The thoracic thickness of 100 patients were measured.Determined the aluminum equivalent of the thoraxes of the infants and young children by comparing the grayscale value and the aluminum step wedge.Another 100 infants and young children of experimental exposure were performed with the aluminum step wedge as a phantom,under AEC control,kV was the only variant to explore the optimal exposure parameters with dose monitor simultaneously.At last,clinical validation was performed.Images quality was compared with x2 test.The radiation dose of two groups was compared with t test.Results The maximum,minimum,average thoracic thickness and their correspondent aluminum equivalent were 13.5 and 2.3 cm,8.0 and 1.4 cm,(10.6 ± 1.3) and 2.0 cm,respectively.The average thoracic thickness of experimental group was (10.1 ± 2.2) cm.The range of entrance surface dose was 0.068-0.056 mGy while the tube voltage range was 55-65 kV.The exposure index range was 0.60-0.74.The visual inspection of aluminum step wedge was from grades 8 to 11 with satisfying image quality at lower radiation.The infant chest X-ray photography exposure parameters formula have been optimized,that was kV =thoracic thickness (cm) × 2 + 38 (constant),mAs (0.8-1.0) with SID =100 cm,without filter grid.Compare to the conventional parameters,the image quality of new method had no significant differences (P ＞ 0.05).The actual average entrance surface dose was (0.048 ± 0.007) mGy,lower than AEC group (0.066 ± 0.008) mGy.The difference was statistically significant (t =16.781,P ＜ 0.001).Conclusions The optimized formula kV =thoracic thickness (cm) ×2 + 38(constant),mAs (0.8-1.0) with SID =100 cm was credible for lowering the radiation exposure with good image quality for clinical diagnosis.

OBJECTIVETo study the protective effect of sufentanil preconditioning against myocardial ischemia-reperfusion (I/R) injury and the role of PI3K/Akt signaling pathway.

METHODSSixty male SD rats weighing 250-350 g were randomly divided into 5 equal groups, namely the sham-operated group, I/R group, sufentanil preconditioning group (Spc group), sufentanil preconditioning +PI3K inhibitor group (Spc+W group), and PI3K inhibitor group (W group). Myocardial I/R model was established by ligation of the anterior descending branch of the left coronary artery for 30 min followed by reperfusion for 120 min. Sufentanil was administered in 3 doses via the femoral vein before the occlusion, each at 1 µg/kg infused within 5 min at a 5-min interval. In Spc+W and W groups, PI3K inhibitor wortmannin (15 µg/kg) was given intravenously 5 min before sufentanil preconditioning and 35 min before ischemia, respectively. Heart rate and mean arterial pressure (MAP) were continuously monitored during I/R. At the end of reperfusion, blood samples were obtained to determine plasma activation of CK-MB and LDH. Acute infarct size was measured by triphenyltetrazolium chloride staining, and the myocardial tissues were obtained to detect the expression of phosphorylated Akt using Western blotting.

RESULTSPhosphorylated Akt expression was significantly up-regulated in I/R and Spc groups as compared with the sham group, and was significantly higher in Spc group than in I/R group. After reperfusion, sufentanil preconditioning significantly decreased myocardial infarct size (P<0.01) and lowered the levels of CK-MB (P<0.01) and LDH (P<0.01) compared with those in the I/R group. The I/R , Spc+W and W groups showed no significant differences in myocardial infarct size or the levels of CK-MB and LDH.

CONCLUSIONThe protective effect of sufentanil preconditioning against myocardium against I/R injury in rats may involve PI3K/Akt signaling pathway activation.

Animals ; Ischemic Preconditioning, Myocardial ; methods ; Male ; Myocardial Reperfusion Injury ; metabolism ; prevention & control ; Proto-Oncogene Proteins c-akt ; metabolism ; Rats ; Signal Transduction ; Sufentanil ; pharmacology

Objective To explore the correlation between postoperative cognitive dysfunction (POCD)and serum level of S100βand NSE in patients undergoing on-pump heart valve replacement. Methods Ninety-eight patients underwent elective heart valve replacement were enrolled and divided into two groups:POCD group (group P)and none POCD group(group NP)by the result of neurocog-nitive testing with MMSE performed preoperatively and on the first postoperative day.Serum S-100βprotein and NSE were measured before operation(T0 ),at the end of operation(T1 ),24 h(T2 )and 48 h(T3 ) postoperatively.Results The incidence of POCD on the first postoperative day was 45 (45.9%).Compared with T0 ,the S100βincreased at T1 in both groups (P <0.05),NSE increased at T1 and T2 in both groups,and NSE increased in group NP at T3 (P <0.05).There was no signifi-cant difference of NSE between groups P and NP.The prolonged recovery time(OR = 1.222,P =0.004)and increased concentration of S100βat the end of operation(OR=1.85,P =0.009)were pre-dictors of POCD on the first postoperative day.Conclusion There was a higher incidence of POCD af-ter on-pump heart valve replacement surgery.The prolonged recovery time and increased concentra-tion of S100βat the end of operation might be predictors of POCD.

An on-line two dimensional liquid chromatographic (2D-LC) method was established by using an ultimate dual gradient liquid chromatography, three chromatographic columns and valve-switching technology to detect 2460A in trichoderma hazianum fermentation liquor. MF C8 column (10 mm×4. 6 mm, 5. 0 μm) was used as purification column and MG C18 column (20 mm×4. 6 mm, 5. 0 μm) was used as enriching column. Methanol and water were used as mobile phase with a gradient elution at a flow rate of 2. 0 mL/min. The sample was separated on the Thermo Hypersil GOLD C18 column (250 mm×4. 6 mm, 5. 0μm) maintained at 40 ℃ using methanol and water. The flow rate was 1. 0 mL/min and 1. 0 mL sample was injected into the 2D-LC system. The detection wavelength was 424 nm. The whole analytical time was less than 60 min. The standard curve was linear over the 2460A concentration range of 0. 0025-10. 0 mg/L(r=0. 9981, n=8). The limit of detection was calculated to be 1 . 2μg/L ( S/N=3 ) and the limit of quantification was calculated to be 2 . 5 μg/L ( S/N=10 ) . The average recoveries varied from 88 . 0% to 104 . 4%.

Objective To study the protective effect of sufentanil preconditioning against myocardial ischemia-reperfusion (I/R) injury and the role of PI3K/Akt signaling pathway. Methods Sixty male SD rats weighing 250-350 g were randomly divided into 5 equal groups, namely the sham-operated group, I/R group, sufentanil preconditioning group (Spc group), sufentanil preconditioning +PI3K inhibitor group (Spc+W group), and PI3K inhibitor group (W group). Myocardial I/R model was established by ligation of the anterior descending branch of the left coronary artery for 30 min followed by reperfusion for 120 min. Sufentanil was administered in 3 doses via the femoral vein before the occlusion, each at 1μg/kg infused within 5 min at a 5-min interval. In Spc+W and W groups, PI3K inhibitor wortmannin (15 μg/kg) was given intravenously 5 min before sufentanil preconditioning and 35 min before ischemia, respectively. Heart rate and mean arterial pressure (MAP) were continuously monitored during I/R. At the end of reperfusion, blood samples were obtained to determine plasma activation of CK-MB and LDH. Acute infarct size was measured by triphenyltetrazolium chloride staining, and the myocardial tissues were obtained to detect the expression of phosphorylated Akt using Western blotting. Results Phosphorylated Akt expression was significantly up-regulated in I/R and Spc groups as compared with the sham group, and was significantly higher in Spc group than in I/R group. After reperfusion, sufentanil preconditioning significantly decreased myocardial infarct size (P<0.01) and lowered the levels of CK-MB (P<0.01) and LDH (P<0.01) compared with those in the I/R group. The I/R , Spc+W and W groups showed no significant differences in myocardial infarct size or the levels of CK-MB and LDH. Conclusion The protective effect of sufentanil preconditioning against myocardium against I/R injury in rats may involve PI3K/Akt signaling pathway activation.

Objective To study the protective effect of sufentanil preconditioning against myocardial ischemia-reperfusion (I/R) injury and the role of PI3K/Akt signaling pathway. Methods Sixty male SD rats weighing 250-350 g were randomly divided into 5 equal groups, namely the sham-operated group, I/R group, sufentanil preconditioning group (Spc group), sufentanil preconditioning +PI3K inhibitor group (Spc+W group), and PI3K inhibitor group (W group). Myocardial I/R model was established by ligation of the anterior descending branch of the left coronary artery for 30 min followed by reperfusion for 120 min. Sufentanil was administered in 3 doses via the femoral vein before the occlusion, each at 1μg/kg infused within 5 min at a 5-min interval. In Spc+W and W groups, PI3K inhibitor wortmannin (15 μg/kg) was given intravenously 5 min before sufentanil preconditioning and 35 min before ischemia, respectively. Heart rate and mean arterial pressure (MAP) were continuously monitored during I/R. At the end of reperfusion, blood samples were obtained to determine plasma activation of CK-MB and LDH. Acute infarct size was measured by triphenyltetrazolium chloride staining, and the myocardial tissues were obtained to detect the expression of phosphorylated Akt using Western blotting. Results Phosphorylated Akt expression was significantly up-regulated in I/R and Spc groups as compared with the sham group, and was significantly higher in Spc group than in I/R group. After reperfusion, sufentanil preconditioning significantly decreased myocardial infarct size (P<0.01) and lowered the levels of CK-MB (P<0.01) and LDH (P<0.01) compared with those in the I/R group. The I/R , Spc+W and W groups showed no significant differences in myocardial infarct size or the levels of CK-MB and LDH. Conclusion The protective effect of sufentanil preconditioning against myocardium against I/R injury in rats may involve PI3K/Akt signaling pathway activation.

Objective To investigate the effect of sufentanil preconditioning on the expression of Toll-like receptor 4 (TLR4) in myocardium during myocardial ischemia-reperfusion (I/R) in rats.Methods Thirty-six male Sprague-Dawley rats,weighing 250-300 g,were randomly divided into 3 groups (n =12 each):sham operation group (group S),I/R group and sufentanil preconditioning group (group SPC).Myocardial ischemia was induced by occlusion of anterior descending branch of left coronary artery for 30 min followed by 2 h of reperfusion.In group SPC,the rats were subjected to 3 consecutive cycles of 5 min sufentanil infusion at 0.2 μg· kg-1 ·min-1 via the femoral vein at 5 min interval before ischemia.In groups S and I/R,the rats were subjected to the equal volume of normal saline instead.HR and mean arterial pressure (MAP) were recorded at 30 min before ischemia (T0),immediately before ischemia (T1),at 30 min of ischemia (T2),and at 30 and 120 min of reperfusion (T3-4).At the end of reperfusion,blood samples were obtained to determine the serum concentration of TNF-α.The rats were then sacrificed and hearts were removed for measurement of myocardial infarct size and expression of TLR4 and NF-κB p65 in myocardium (using Western blot).Results There was no significant difference in HR among the three groups (P ＞ 0.05).Compared with group S,MAP was significantly decreased at T2-4,the serum concentration of TNF-α was increased,and the expression of TLR4 and NF-kB p65 protein in myocardium was upregulated in groups I/R and SPC (P ＜ 0.01).Compared with group I/R,no significant difference was found in MAP at all time points (P ＞ 0.05),and myocardial infarct size was significantly decreased,serum concentrations of TNF-α were decreased,and the expression of TLR4 and NF-κB p65 protein in myocardium was down-regulated in group SPC (P ＜ 0.01).Conclusion The mechanism by which sufentanil preconditioning alleviates myocardial I/R injury may be related to down-regulation of TLR4 expression in rat myocardium.

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.