Objective To develop SPE - HPLC method for the assay of paeonol in Liuwei Dihuang Pill (LDP). Methods Separation was performed on a Hypersil ODS(2) column (250 mm?4. 6 mm, 5?m) . The mobile phase consist of methanol - 2 % acetic acid (55 :45, v/v) . The flow rate was 1 mL/min. The UV detection was set at 275 nm. Results Paeonol had a good linear relation in the range of 0. 03998 ~ 0. 7996 ug, the average recovery of honeyed bolus of LDP was 102. 9 % and the RSD was 1.2%; the average recovery of concentrated bolus of LDP was 101. 5 % and the RSD was 1.8%. Conclusion The method is simple, sensitive and rapid and it can be used to determine the paeonol content of LDP.

A novel citrinin derivative, penicitrinol L (1), along with two known analogues, penidicitrinin B (2) and pennicitrinone A (3) were isolated from the marine-source fungus Penicillium citrinum. The structure of the new compound was elucidated by spectroscopic methods including one and two-dimensional NMR as well as high-resolution mass spectrometric analysis. Furthermore, compound 1 showed modest cytotoxic activity against HL-60 cell line and compound 3 showed weak cytotoxic activity against A375 cell line.

Objective To validate the analytical performance of three Cys C reagents with particle-enhanced turbidimetric immunoassay(PETIA) method used on the automatic biochemistry analyzer for preliminary clinical application.Methods The performance of three Cys C reagents (labeled as A, B, C) with PETIA method from Shanghai Jing Yuan Co., Beijing Leadman Co. and Beijing Jiuqiang Co. on OlympusAU2700 automatic biochemistry analyzer were assessed.According to the standard of CLSI EP6-A, EP15-A and EP7-P, the precision, linearity range, disturbance (bilirubin, hemoglobin, chyle) were assessed, and compared with those of Cys C reagent based on particle-enhanced nephelometric immunoassay(PENIA) from Dade Behring Co.. The reference ranges for Cys C in serum of 120 healthy individual were evaluated.Results The within-run CVs of the three reagents (A, B and C) were 3.08%-3.2%, 2.3%-4.15% and 1.38%-1.53% respectively.The total CV in A, B and C were 3.29%-3.44%, 2.65%-5.18% and 1.67%-1.69% respectively, lower than the stated.Limits of quantitative determination (LOQ) of the three reagents were 0.41, 0.23 and 0.07 mg/L, basically meeting the testing requirement.The linearity range was 0.22-7.26 mg/L(r=0.996), 0.20-7.72 mg/L(r=0.999)and 0.20-7.62 mg/L(r=0.997)in the three reagents, which demonstrated a sound linear correlation. For interference tests, no remarkable interference (<±10%) of reagent C was detected when bilirubin≤684 μmol/L, hemoglobin≤9.7 g/L and Chyle turbidity≤6 200 FTU; and no significant interference of reagent B was found when bilirubin≤684 μmol/L, hemoglobin≤6.79 g/L and Chyle turbidity≤6 200 FTU; when bilirubin≤684 μmol/L, hemoglobin≤4.85 g/L and Chyle turbidity≤1 240 FTU reagent A was not interfered significantly. The comparison afte and before the high-speed centrifugation reveals that the average percentage of bias for reagents A, B, C measured Cys C in chylous serum samples of patients was -8.31%, 1.52%, 1.32%, respectively.In method comparison tests, the regression equations of the three reagents compared with Dade Behring PENIA Cys C reagent were as follows:Y=0.787X+0.492 (R2=0.976), Y=1.098X+0.137 (R2=0.982) and Y=1.037X+0.249 (R2=0.996), respectively. Agreement rates of the high Cys C in reagent A, B, C and Dade Behring Cys C reagent were 80% (Kappa=0.615,P=0.000), 100% (Kappa=1.000,P=0.000), 91.2% (Kappa=0.824,P=0.000); While for reference range of preliminary clinical assessment, diagnosis coincidence rate of reagent A increased to 98.8% (Kappa=0.974,P=0.000). Conclusions When used in automatic biochemical analyzer, the three Cys C reagent with PETIA showed high precision,sensitivity, and sound correlation with Dade Behring PENIA reagents.The three reagents are all able to meet clinical test requirements, nevertheless, anti-interference capability were diffierent and the reference range should be further validated.

Objective:To investigate the effect of enhanced external counterpulsation(EECP)on heart failure with preserved ejection fraction(HFpEF)and hemodynamics in elderly patients.Methods:Clinical data of 66 elderly HFpEF patients admitted to the enhanced external counterpulsation center of our hospital from January 2018 to December 2019 were retrospectively analyzed.The primary assessment parameter was the six-minute walk distance, and the secondary parameters included the Minnesota Living with Heart Failure Questionnaire(MLHFQ). Noninvasive hemodynamic parameters including the cardiac index(CI), stroke volume(SV), isovolumic relaxation period(A2-mitral valve opening, A2-O), pulmonary capillary wedge pressure(PCWP)and total peripheral resistance(TPR)were monitored and mean arterial blood pressure(MAP)was calculated.Results:After EECP treatment, the six-minute walk distance increased(372±87 m vs.341±85 m, P<0.05), the score of MLHFQ decreased(47.0±16.0 vs.50.0±17.0, P<0.05), CI increased(2.8±0.7)L·min -1·m -2vs.(2.6±0.6)L·min -1·m -2( P<0.05), SV, PCWP and A2-O did not show significant change(73.3±16.4 ml vs.71.5±17.1 ml, 10.0±3.3 mmHg vs.11.0±3.6 mmHg, 1 mmHg=0.133 kPa, 98.0±29.5 ms vs.91.0±29.1 ms, P>0.05), TPR decreased(1 719.0±427.0 DS/cm 5vs.1 821.0±508.0 DS/cm 5, P<0.05), and there was no significant change in MAP(96.9±10.7 mmHg vs.98.8±13.1 mmHg, P>0.05), compared with pre-EECP treatment levels.Compared with patients without hypertension, MAP decreased in patients with hypertension(14 cases), when stratified by the initial MAP( P<0.05). Conclusions:For elderly patients with HFpEF, EECP can increase the six-minute walk distance, improve the quality of life and hemodynamic parameters, and is a safe adjuvant treatment.

Objective To investigate the reference values and Z scores regression equations of newborn undergoing echocardiography. Methods Two hundred and eighty-eight newborns (aged 0-28 days) of Shenzhen Children′s Hospital underwent echocardiography examination, including M-mode, two-dimensional (2D) and real-time three-dimensional (3D) echocardiography, color Doppler lfow imaging (CDFI) and tissue Doppler imaging. The correlation between echocardiography results and weight were analyzed and Z scores were calculated. Results The normal values of right ventricular diameter (RV) and left ventricular end-diastolic diameter (LVEDD) measured by M-mode, the mitral annulus diameter in four chamber view (MV-D1), mitral annulus diameter in two chamber view (MV-D2), mitral annulus diameter in longitudinal view (MV-D3), aortic ring diameter (ARD), aortic sinus diameter (ASD), ascending aorta diameter (AAO), transverse aorta diameter (TA), aortic isthmus diameter (AI), aorta diaphragm diameter (AO-Dia), tricuspid annulus diameter in four chamber view (TV-D1), tricuspid annulus diameter in right ventricular inlfow tract view (TV-D2), right ventricular outlfow tract diameter (RVOT), pulmonary valve diameter (PVD) and main pulmonary artery diameter (PA) measured by 2D echocardiography and the normal values of mitral valve inflow Doppler component during early diastole (MV-E), mitral valve inlfow Doppler component during atrial contraction (MV-A), tricuspid valve inlfow Doppler component during early diastole (TV-E), tricuspid valve inflow Doppler component during atrial contraction (TV-A), aortic valve peak velocity (AV-max), aortic valve velocity-time integral (AV-VTI), pulmonary valve peak velocity (PV-max), pulmonary valve velocity-time integral (PV-VTI) measured by pulse Doppler, the mitral annular tissue Doppler component during systole (MV-s′), mitral annular tissue Doppler component during early diastole (MV-e′), mitral annular tissue Doppler component during atrial contraction (MV-a′), tricuspid annular tissue Doppler component during systole (TV-s′), tricuspid annular tissue Doppler component during early diastole (TV-e′), tricuspid annular tissue Doppler component during atrial contraction (TV-a′), interventricular septum annular tissue Doppler component during systole (IVS-s′), interventricular septum annular tissue Doppler component during early diastole (IVS-e′), interventricular septum annular tissue Doppler component during atrial contraction (IVS-a′) measured by tissue Doppler, the normal values of left atrial volume (LAV), left ventricular end-systolic volume (LVEDV), stroke volume (SV) and cardiac output (CO) measured by bi-plane method and the normal values of LVEDV, SV and CO measured by real-time tri-plane method, together with the normal values of left ventricular (LV) mass, left ventricular mass index [LV mass/BSA, LV mass/H2.7, body surface area (BSA) and height (H)], all showed nonlinear positive correlations with body weight (all P<0.01). The values of MV-E/A, PV-E/A, MV-e′/a′, TV-e′/a′, IVS-e′/a′, MV-E/IVS-e′, LV mass/LVEDV and left ventricular ejection fraction (LVEF) showed no correlations with body weight (all P>0.05). Except for RV, MV-D1, MV-D2, MV-D3, TV-D1, TV-E, MV-s′, IVS-a′, TV-s′and TV-e′, all R2 obtained by nonlinear regression method (lnY=a+bX+cX2+dX3) were larger than those obtained by linear regression method (Y=a+bX). The Z score showed a normal distribution and no correlation with body weight. Conclusions The normal reference values of newborn undergoing echocardiography reflect the variation in weight. The Z scores can be obtained by the predicted nonlinear regression equations and show standard normal distribution. The echocardiography normal reference values have important significance for the diagnosis and treatment of neonatal heart disease.

Objective To investigate the value of ultrasound in tethered cord syndrome (TCS) in infants. Methods From December 2005 to July 2013, 25 cases TCS were confirmed by surgery in Shenzhen children's hospital. The ultrasonogram and MRI of 25 infants were analysed retrospectively. The diagnostic accuracy of ultra-sound and its clinical significance were evaluated. Results In 25 cases of TCS confirmed by surgery, the coincidence rate of conus level detected by ultrasound [96%(23/24)] was lower than that by MRI (100%(25/25)). Ultrasound showed reduced spinal cord vibration in 4 cases, disappeared spinal cord vibration in 21 cases. Two cases of reduced spinal cord vibration detected by ultrasound were confirmed as disappeared spinal cord vibration by surgery. The coincidence rate of disappeared spinal cord vibration detected by ultrasound was 91%(21/23). The sacral spinal seg-ments form showed by ultrasound were entirely consistent with those of MRI, including 4 cases of enlarged spinal cord, 13 cases of spinal cord without enlargement and 8 cases of spinal cord ended with rat caudate. Twenty-five cases of TCS had malformations:7 cases meningocele (3 cases combined lipoma), 14 cases myelomeningocele (5 cases combined lipoma, 1 case combined hydromyelia), 3 cases spinal canal-epidermis fistula (all combined lipoma) and 1 case solitary lipoma. Compared with the operation findings, ultrasound misdiagnosed 2 cases of myelomeningo-cele as meningocele, missed 1 of case lipoma which combined with meningocele. MRI missed two cases of spinal canal-epidermis fistula. Conclusions Infantile spinal ultrasound examination can accurately locate the position of conus, accurately display the spinal cord. Compared with MRI, ultrasound examination can real-time visually display spinal cord vibration and help to diagnose tethered cord. Ultrasound examination are convenient, repeatable opera-tion with low cost, therefore it can be used as the preferred screening method to diagnose of tethered spinal cord.

ObjectiveTo investigate the impacts of different serum creatinine detection methods,including Jaffe and enzymatic methods,on the efficacy of different GFR estimation equations in CKD patients in China.MethodsrGFR of 176 patients with CKD were determined by dual plasma sample method 99mTc-diethylenetriamine pentaacetic acid (99mTc-DTPA) plasma clearance rate.Serum creatinine was detected with four kinds of creatinine reagents from different manufacturers.Cockcroft-Gault Equation corrected for body surface area (CG/BSA),simplified Modification of Diet in Renal Disease (MDRD) Study equation,IDMS-traceable MDRD equation,CKD epidemiology collaborative research (CKD-EPI) equation and two Chinese simplified MDRD equation (project group equation 1,2) were applied to calculate estimated GFR (eGFR)respectively.eGFRwerecomparedwithrGFRforthecorrelation, deviation, precisionand30％ accuracy.ResultsThe mean rGFR of 176 patients with CKD,was [ 40.70 ( 19.41 -84.35 ) ] ml · min- 1 ·( 1.73 m2 ) -1.For all GFR estimation equations,there were significant differences in eGFR results between enzymatic method and Jaffe method,when analyzed by the Wilcoxon signed-rank test.eGFR results assessed by two enzymatic creatinine detection systems showed no significant difference,while eGFR results analyzed by two Jaffe detection system were significantly different.The intraclass correlation coefficient (ICC) of eGFR and rGFR ranged from 0.879 to 0.923 by Jaffe method,while from 0.925 to 0.946 by enzymatic creatinine method.ICC and Pearson correlation analysis revealed a significant correlation between eGFR and rGFR,and the correlation was better when using enzymatic method.Bland-Altman plots indicated that large deviation occurred in the high value area of GFR using various equations.However,deviation with the enzymatic creatinine method was smaller than that with the Jaffe method. When rGFR ≥ 60 ml · min- 1 ·(1.73 m2) -1,the 30％ accuracy of eGFR using enzymatic creatinine method for all six equations was between 68.3％ and 90.0％,while it was between 41％ and 75％ when using Jaffe method. The 30％ accuracy of eGFR using enzymatic creatinine method was significantly higher than that using picric acid method for these equations except for the project group equation 1.When rGFR ＜60 ml · min -1 · ( 1.73 m2 ) -1,the 30％accuracy of eGFR using both methods was between 39.7％ -49.1％,40.5％ -52.6％respectively,and the difference of data showed no statistical significance.For the same equation,there was a significant differernce in 30％ accuracy of eGFR between two enzymatic creatinine detection systems,while there was no significant differernce between two Jaffe creatinine detection systems.ConclusionsA significant difference was demonstrated in the same GFR evaluation equation using two different creatinine detection methods (Jaffe method and enzymatic method).The correlation between rGFR and eGFR,the degree of deviation,and accuracy of eGFR results assessed by enzymatic creatinine method were better than those by Jaffe method.The eGFR results assessed by different enzymatic detection systems revealed no significant difference.

Objective To investigate the clinical value of coronary artery Z-scores on echocardiography in diagnosing coronary artery abnormalities. Methods The echocardiography results of 612 patients with Kawasaki disease (KD) at the acute and recovery phase were retrospectively studied. Coronary artery luminal diameters were converted to body-surface-area-adjusted Z-scores. According to coronary Z-scores classiifcation, all the subjects were divided to four groups:415 cases with no dilation (ND), 133 cases with small coronary artery abnormalities (SCAAs), 47 cases with large coronary artery abnormalities (LCAAs), and 17 cases with giant coronary artery abnormalities (GCAAs). Clinical features (gender, age, typical clinical manifestations, fever duration) and laboratory results (CRP, ESR, WBC, PLT) were compared among all the four groups. Coronary artery diameters and the Z-scores were compared between acute and convalescence phase. Results Along with the increase of coronary Z-score, fever duration was prolonged [ND group:(7.75±3.12) d, SCAAs group (8.50±4.12) d, LCAAs group: (8.57±3.58) d, GCAAs group: (11.88±4.33) d, F=22.375, P<0.05]. With coronary Z-score increasing, PLT also increased (F=22.029, P=0.000), and the highest PLT was observed in GCAAs group. There were no significant differences in the CRP, ESR and WBC among all the four groups (F=0.236, 1.116, 0.121, all P>0.05). No significant different coronary diameters were found in ND cases between recovery and acute phase [(2.24±0.34) mm vs (2.33±0.36) mm, t=1.926, P > 0.05]. But there were significant difference in the coronary Z-scores of ND patients between recovery and acute phase (0.41±0.82 vs 1.17±0.75, t=8.332, P < 0.05). The coronary Z-scores in SCAAs group (1.32±0.89 vs 3.40±0.62, t=11.073, P < 0.05), LCAAs group (3.12±2.27 vs 6.20±1.28, t=4.579, P<0.05) and GCAAs group (11.88±6.77 vs 20.4±9.70, t=3.480, P<0.05) at recovery phase were smaller than values at acute phase. Conclusions The KD coronary Z-scores are the body-surface-area-adjusted standard value, and not subject to the influence of children growth and development. Therefore, it may accurately evaluate the severity of coronary artery abnormalities and its recovery process. Accurate quantitative of the coronary artery luminal dimensions is important in KD clinical management and prognosis prediction.

Objective:To investigate the influence of ferri ion on the growth, reproduction and energy metabolism of Leptospira interrogans ( L. interrogans), and to identify whether the LA_2690 and LA_3598 gene products functioned as ferritin and ferroxidase. Methods:Petroff-Hausser counting method was used to analyze the influence of ferri ion deficiency on the growth and reproduction of L. interrogans serogroup Icterohaemorrhagiae serovar Lai strain 56601 in EMJH medium. Spectrophotometry and Chemiluminescence method was used to detect whether ferri ion deficiency inhibited the synthesis of DNA and ATP in L. interrogans. The structures and functions of L. interrogans LA_2690 and LA_3598 genes were analyzed using bioinformatic softwares. Prokaryotic expression systems for LA_2690 and LA_3598 genes were established and the target proteins, rLep2690 and rLep3598, were extracted by Ni-NTA affinity chromatography. The ferroxidase activity of rLep2690 and rLep3598 was detected by spectrophotometry. After L. interrogans strain 56601 was used to infect human umbilical vein endothelial cells (HUVEC) and monocytes (THP-1), changes in the expression of LA_2690 and LA_3598 genes at transcription level were detected using real-time fluorescence quantitative RT-PCR (qRT-PCR). Results:In the ferri ion-absent EMJH medium, the growth and reproduction of L. interrogans as well as the DNA and ATP synthesis levels were significantly decreased ( P<0.05). The products of LA_2690 and LA_3598 genes were predicted as bacterioferritin (Bfr) and DNA-binding ferritin containing ferroxidase diiron centers, but the latter lacked the heme-binding site and ferroxidase core. The prokaryotic expression systems for LA_2690 and LA_3598 genes could efficiently express the target recombinant proteins. Both the purified rLep2690 and rLep3598 showed a single band on SDS-PAGE. The ferroxidase activity of rLep2690 and rLep3598 was 1 238.619 U/L and 60.052 U/L, respectively. The expression of LA_2690 and LA_3598 genes of L. interrogans at mRNA level was significantly elevated during infection of the two types of cells ( P<0.05). Conclusions:Ferri ion participates in the growth and reproduction of L. interrogans as well as the synthesis of DNA and ATP. LA_2690 and LA_3598 genes were essential for L. interrogans to infect cells, and the product of LA_2690 gene possessed a stronger ferroxidase activity.

Objective:To compare the diagnostic accuracy of lung ultrasonography and radiography in detection of acute traumatic intrathoracic injuries in children.Methods:A retrospective analysis was performed in 46 cases of children with chest trauma in Shenzhen Children′s Hospital from January 2017 to January 2021. The diagnostic efficiency of lung ultrasound and radiography in children with acute traumatic hemopneumothorax and lung contusion were compared. Computed tomography scan was used as gold standard.Results:The sensitivity and specificity of lung ultrasound were 0.79 and 0.98 for pneumothorax, 0.86 and 0.75 for hemothorax, 0.86 and 0.80 for pulmonary contusion, respectively. Area under the ROC curve of ultrasonography in detection of pneumothorax, hemothorax, and pulmonary contusion was 0.889 (95% CI=0.798-0.979, P<0.001), 0.804 (95% CI=0.707-0.901, P<0.001), and 0.831 (95% CI=0.623-1.000, P=0.013), respectively. Area under the ROC curve of radiography was 0.674 (95% CI=0.544-0.803, P=0.008) for detection of pneumothorax, 0.645 (95% CI=0.517-0.772, P=0.026) for hemothorax, and 0.547 (95% CI=0.289-0.805, P=0.724) for pulmonary contusion. Comparison of area under the ROC curve declared the significant superiority of ultrasonography in detection of pneumothorax, hemothorax and pulmonary contusion (all P<0.05). Conclusions:Lung ultrasound has higher sensitivity and accuracy than chest radiography in the initial evaluation of chest trauma. For children with acute chest trauma, lung ultrasound should be the first choice.