Background: Twenty-four-hour urinary free cortisol (UFC) measurement is the initial diagnostic test for Cushing’s syndrome (CS). We compared UFC determination by both direct and extraction immunoassays using Abbott Architect, Siemens Atellica Solution, and Beckman DxI800 with liquid chromatography-tandem mass spectrometry (LC-MS/MS). In addition, we evaluated the value of 24-hr UFC measured by six methods for diagnosing CS. Methods: Residual 24-hr urine samples of 94 CS and 246 non-CS patients were collected.A laboratory-developed LC-MS/MS method was used as reference. UFC was measured by direct assays (D) using Abbott, Siemens, and Beckman platforms and by extraction assays (E) using Siemens and Beckman platforms. Method was compared using Passing–Bablok regression and Bland–Altman plot analyses. Cut-off values for the six assays and corresponding sensitivities and specificities were calculated by ROC analysis. Results: Abbott-D, Beckman-E, Siemens-E, and Siemens-D showed strong correlations with LC-MS/MS (Spearman coefficient r = 0.965, 0.922, 0.922, and 0.897, respectively), while Beckman-D showed weaker correlation (r = 0.755). All immunoassays showed proportionally positive bias. The areas under the curve were 0.975 for Abbott-D, 0.972 for LCMS/MS, 0.966 for Siemens-E, 0.948 for Siemens-D, 0.955 for Beckman-E, and 0.877 for Beckman-D. The cut-off values varied significantly (154.8–1,321.5 nmol/24 hrs). Assay sensitivity and specificity ranged from 76.1% to 93.2% and from 93.0% to 97.1%, respectively. Conclusions Commercially available immunoassays for measuring UFC show different levels of analytical consistency compared to LC-MS/MS. Abbott-D, Siemens-E, and Beckman-E have high diagnostic accuracy for CS.

Objective To investigate the mechanism of astragaloside Ⅰ,the active constituent of milkvetch root,in inhibiting podocyte injury and improving diabetic kidney disease.Methods According to the body weight,60 male db/db mice were randomly divided into the model group,astragaloside Ⅰ low-dose group(10 mg/kg),astragaloside Ⅰ medium-dose group(20 mg/kg),astragaloside Ⅰ high-dose group(40 mg/kg),and valsartan group(10mg/kg),with 12 mice per group.Twelve db/db littermate control db/m mice were used as the control group.The drug was administered by gavage for 8 weeks.Transmission electron microscope was used to observe the ultrastructure of the kidney;immunohistochemistry and Western blotting were used to detect the expression of nephrotic protein(nephrin),a marker of renal podocytes;enzyme-linked immunosorbent assay was used to detect the contents of interleukin-1β(IL-1β)and interleukin-18(IL-18)in the serum of mice;Western blotting was used to detect the protein expressions of NOD-like receptor thermoprotein domain-related protein 3(NLRP3),cysteinyl aspartate specific proteinase 1(Caspase-1),and Gasdermin D(GSDMD)in kidney tissue.Results Compared with the control group,the glomeruli of the model group showed obvious podocyte loss and foot process fusion;the protein expression of nephrin was decreased(P＜0.05);the contents of IL-1 β and IL-18 in serum were increased(P＜0.05);the protein expressions of NLRP3,Cleaved-Caspase-1,and GSDMD-N were increased(P＜0.05).Compared with the model group,the renal pathological damage in the astragaloside Ⅰ administration groups were alleviated;the protein expression of nephrin was increased(P＜0.05);the contents of IL-1β and IL-18 in serum were decreased(P＜0.05);the protein expressions of NLRP3,Cleaved-Caspase-1,and GSDMD-N were decreased(P＜0.05).Conclusion Astragaloside Ⅰ may play a role in intervening diabetic kidney disease by inhibiting pyroptosis and improving podocyte injury.

OBJECTIVE To investigate the effect of ropivacaine combined with dexmedetomidine on postoperative analgesia in women undergoing cesarean section， and to explore the feasibility of the opioid-free analgesia mode after cesarean section under spinal-epidural anesthesia. METHODS Totally 80 women undergoing cesarean section were randomly divided into observation group （ropivacaine combined with dexmedetomidine for analgesia） and control group （ropivacaine combined with opioid drug sufentanil for analgesia） ， with 40 cases in each group. The exercise and rest score in visual analogue scale （VAS） within 48 hours after operation， the use of analgesia pump （the time of first analgesia pump pressing， the times of analgesia pump pressing within 24 hours and 48 hours after operation）， the time of block （the onset time of spinal anesthesia sensory block， the time to the highest level of spinal anesthesia sensory block， the time of sensory recovery and the time of movement recovery） ， the time of prognosis （the time of gastrointestinal ventilation recovery， the time of getting out of bed and the hospitalization time）， and the incidence of adverse events were compared in 2 groups. RESULTS Finally， 64 parturients （32 in the observation group and 32 in the control group） were involved in the analysis. Compared with the control group， the recovery time of sensation and movement were significantly prolonged， the ventilation time was significantly shortened， and the incidence of nausea， vomiting and abdominal distension was significantly decreased in the observation group （P＜0.05） . There was no significant difference in the other indexes between the two groups （P＞0.05）. CONCLUSIONS Ropivacaine combined with dexmedetomidine under spinal-epidural anesthesia could provide similar analgesic effect as combined with opioids drug sufentanil， shorten the time of gastrointestinal ventilation recovery， and reduce the incidence of nausea，vomiting and abdominal distension， with no increased risk of low blood pressure or urinary retention.

Objective:To investigate associations between clinicopathological characteristics and mutations in susceptibility genes in cutaneous melanoma (CMM) .Methods:A total of 94 patients with confirmed CMM were collected from People′s Hospital of Xinjiang Uygur Autonomous Region from January to December in 2019, and their clinical and histopathological characteristics were retrospectively analyzed. In 48 paraffin-embedded melanoma tissue specimens, Sanger sequencing was performed to detect mutations in the BRAF, NRAS, c-KIT genes and the promoter region of human telomerase reverse transcriptase (hTERT) gene, and the association between gene mutations and clinicopathological characteristics was analyzed. Measurement data were compared using t test, and enumeration data were compared using chi-square test or Fisher′s exact test. Results:Among the 94 patients with CMM, there were 46 (48.9%) males and 48 (51.1%) females, with the age being 58.5 ± 16.0 years; 41 (43.6%) patients were of Han nationality, and 53 (56.4%) were of ethnic minorities. Skin lesions were located at the acral sites in 50 (53.2%) patients, including 27 (28.7%) of Han nationality; non-acral skin lesions occurred in 44 (46.8%) , including 14 (31.8%) of Han nationality; there was a significant difference in the nationality distribution between the acral CMM group and non-acral CMM group ( χ2 = 5.25, P = 0.022) . Histopathological examination showed CMM of Clark grades Ⅳ or Ⅴ in 41 (43.6%) cases, ulcers in 52 (55.3%) cases, and lymph node metastasis in 32 (34.04%) cases at the first clinic visit. Gene sequencing revealed BRAF gene mutations in 11 (22.9%) of 48 cases, including c.1799 T>A (p.V600E) , c.1790 T>A (p.L597Q) and c.1394 C>T (p.S465F) ; NRAS gene mutation c.182 A>G (p.Q61R) was identified in 5 (10.4%) cases; c-KIT gene mutations were identified in 6 (12.5%) cases, including c.1727 T>C (p.L576P) and c.1669 T>C (p.W557R) ; mutations in the promoter region of hTERT gene were identified in 7 (14.6%) cases, including 4 cases with a mutation at 124 bp upstream of the ATG start codon (C228T) and 3 cases with a mutation at 146 bp upstream of the ATG start codon (C250T) . Among 26 patients aged < 60 years, BRAF gene mutations were found in 9, and the incidence of BRAF gene mutations was significantly higher in the patients aged < 60 years than in those aged ≥ 60 years (2/22, P < 0.05) , but significantly lower in the patients with acral CMM (3/27) than in those with non-acral CMM (8/21, P < 0.05) ; the incidences of the NRAS, c-KIT and hTERT gene mutations were all significantly higher in the patients with lymph node metastases (3/10, 4/10, 4/10, respectively) than in those without (2/38, 2/38, 3/38, respectively, all P < 0.05) . Conclusion:CMM lesion locations significantly differed among different ethnic groups; the BRAF gene mutation was associated with the age of patients and lesion locations of CMM; NRAS, c-KIT gene mutations and hTERT promoter mutations were closely related to lymph node metastasis.

Objective To investigate the relationship between the mechanism underlying hydrogeninduced reduction of sepsis-associated encephalopathy (SAE) and phenotypic transformation of hippocampal microglias in mice.Methods Eighty-eight adult male ICR mice,aged 6-8 weeks,weighing 20-25 g,were divided into 4 groups (n =22 each) using a random number table method:sham operation group (group Sham),sham operation plus hydrogen group (group Sham+H2),SAE group and SAE plus hydrogen group (group SAE + H2).Sepsis was induced by cecal ligation and puncture (CLP) in anesthetized mice.Sham and Sham+H2 groups only underwent simple laparotomy.Sham+H2 and SAE+H2 groups inhaled air containing 2％ hydrogen for 1 h starting from 1 and 6 h after CLP,respectively.Mice were sacrificed at 24 h after CLP,and hippocampi were isolated for determination of the levels of tumor necrosis factor (TNF-α),interleukin-6 (IL-6),transforming growth factor-β (TGF-β) and IL-10 (by enzyme-linked immunosorbent assay) and expression of inducible nitric oxide synthase (iNOS) and argininase-1 (Arg-1) (by Western blot).Morris water maze test was performed on 10 mice in each group at days 4-8 after CLP.PResults Compared with group Sham,the levels of TNF-α,IL-6,TGF-β and IL-10 were significantly increased,the expression of iNOS and Arg-1 was up-regulated,the escape latency was prolonged,and the rate of time spent in the target quadrant and the number of crossing the original platform were reduced in SAE and SAE+H2 groups (P＜0.05).Compared with group SAE,the levels of TNF-α and IL-6 were significantly decreased,the expression of iNOS was down-regulated,the expression of TGF-β,IL-10 and Arg-1 was up-regulated,the escape latency was shortened,and the rate of time spent in the target quadrant and the number of crossing the original platform were increased in group SAE+H2 (P＜0.05).Conclusion Hydrogen can promote phenotypic transformation of hippocampal microglias from M1 to M2 and reduce SAE in mice.

Objective: To evaluate the effect of anesthesia management based on regional cerebral oxygen saturation (rSO₂)-bispectral index (BIS)-goal-directed hemodynamic multi-modal monitoring on acute kidney injury (AKI) after cardiac valve replacement. Methods: A total of 238 patients of both sexes, aged 18-75 yr, weighing 45-95 kg, of American Society of Anesthesiologists physical status Ⅲ or Ⅳ (New York Heart Association class Ⅱ or Ⅲ ), undergoing elective cardiac valve replacement, were assigned into routine experience group (group C, n=122) and multi-mode monitoring group (group M, n=116) using a random number table method.After admission to the operating room, the invasive blood pressure of radial artery, central venous pressure, electrocardiogram, SpO₂ and end-tidal pressure of carbon dioxide were recorded in group C, and rSO₂, BIS, stroke volume variation, cardiac index and stroke volume index were monitored based on the monitoring in group C. The patients were tracheally intubated when BIS value was about 50.BIS value was maintained between 45 and 55, rSO₂ was maintained not less than 20% of the baseline value or the absolute value not less than 55%, and fluctuation in mean arterial pressure did not exceed 20% of the baseline value, and stroke volume index was maintained >25 ml/m² and cardiac index>2.5 L·min^-1 ·m^-2, maintaining not lower than the baseline value before induction in special conditions.The intraoperative volume of fluid infused, infusion of allogeneic blood, consumption of propofol, time of cardiopulmonary bypass, aortic cross-clamping time, anesthesia time, operation time, occurrence of AKI within 48 h after operation, renal replacement therapy, extubation time, duration of intensive care unit stay, postoperative length of hospital stay, development of other serious postoperative complications and fatality rate at day 30 after operation were recorded. Results: Compared with group C, the incidence of AKI was significantly decreased, the volume of plasma infused and consumption of propofol were reduced, the extubation time, duration of intensive care unit stay and postoperative length of hospital stay were shortened (P<0.05), and no significant change was found in the rate of renal replacement therapy, constituent ratio of AKI degree, incidence of other serious postoperative complications or mortality rate at day 30 after operation in group M (P>0.05). Conclusion Anesthesia management based on rSO₂-BIS-goal-directed hemodynamic multi-modal monitoring can decrease the occurrence of AKI after cardiac valve replacement.

Objective: To compare the differences and similarities in emotional and behavioral problems among children with developmental dyslexia (DD), attention deficit hyperactivity disorder (ADHD) or both, aiming to provide basis for targeted intervention. Methods: Strengths and Difficulties Questionnaire (SDQ) was used to investigate emotional and behavioral problems among four groups of children (DD=32, ADHD=32, DD+ADHD=14, TD=32). Hierarchical regression analysis was recruited to investigate the associations of emotional and behavioral problems with spelling impairment or ADHD symptoms. Results: Except for peer problems, emotional and behavioral problems varied in the four groups [prosocial DD: (641±2.75), ADHD: (5.53±2.49), combined: (7.07±2.37), control: (7.72±2.19), F=4.42, P<0.05; emotional problems DD: (1.81±1.94), ADHD: (3.66±1.66), combined: (3.21±2.33), control: (2.03±1.99), F=6.36， P<0.05; conduct problems DD: (2.31±1.23), ADHD: (3.66±1.77), combined: (2.64±1.98), control: (1.97±1.31), F=7.24, P<0.05; difficulties score DD: (6.94±2.96), ADHD: (10.28±3.90), combined: (9.07±3.17), control: (6.06±3.94), F=8.82, P<0.05 ], most serious in ADHD group. In addition, children in DD group scored higher than those in normal group in terms of prosocial behavior. Children in the combined group showed higher total scores of peer problems and difficult score than those in the normal group. Emotional symptoms were more serious among children in combined group than those in the DD group. Compared with the combined group, behavioral problems in ADHD group were more common. Hierarchical regression analysis indicated that occurrence of emotional and behavioral problems in combined group was mostly mediated by hyperactivity symptoms. Conclusion Compared with DD group, children with ADHD combined or not combined with DD show more emotional and behavioral problems. Emotional and behavioral problems are notably observed among those with hyperactivity symptoms than DD.

Objective To establish the Nifedipine-induced liver cell damage model,and to investigate the cellular or molecular mechanism for children's liver cell damage.Methods The HepG2 cells were utilized to establish liver cell damage models.The optimal concentration and the optimal pretreatment time of Nifedipine-induced liver cell injury were confirmed.Western blot and real-time PCR(RT-PCR)were used to check the alteration in proteins and mRNAs level of the liver function-associated classic markers,which contained alkaline phosphatase(ALP),aspartate amino transferase(AST),glutamyl transpeptidase(γ-GT)and alanine aminotransferase(ALT).Additionally,flow cytometry(FCM),colony formation assay(CFA)and cell wound healing assay(CWHA)were utilized to check the effect of Nifedipine on the cell cycle progression and proliferation of HepG2.Results (1)The optimal concentration of Nifedipine was 20 mg/L and the optimal treatment period was 21 days for liver damage.(2)Western blot:intracellular ALT protein content after Nifedipine group was less than that of control group,while the protein levels of AST,γ-GT and ALP in the culture medium after Nifedipine addition(3.55 ± 0.05,4.91 ± 0.055,3.51 ± 0.05,3.08 ± 0.08) were higher than those of control group(0.96 ± 0.02,1.03 ± 0.02,1.00 ± 0.05,0.90 ± 0.13),and the differences were all significant(t = -85.695,-117.582,-47.371,-33.260,all P<0.05).(3)The findings of RT-PCR showed that the mRNA levels of intracellular ALT,γ-GT and ALP in Nifedipine group(0.26 ± 0.02,0.05 ± 0.04, 0.05 ± 0.02)were lower than those of control group(1.13 ± 0.21,0.94 ± 0.10,1.03 ± 0.06),and the differences were all significant(t=7.233,127.436,25.687,all P<0.05).However,the mRNAs levels in purified culture me-dium in Nifedipine group(5.95 ± 0.05,3.13 ± 0.10,3.32 ± 0.08)were higher than those in control group(1.01 ± 0.08,1.00 ± 0.05,1.00 ± 0.05),and the differences were all significant(t= -92.339,-31.250,-43.007,all P<0.05).(4)The portion of G0/G1 phase in Nifedipine group[(84.09 ± 0.43)%]was more than that of control group[(30.93 ± 0.32)%],which had statistical significance(t=173.084,P=0.000).(5)In contrast with control group,the colony formation of cells in Nifedipine group declined from(97.10 ± 1.17)% to(38.56 ± 1.51)%(t=92.088,P=0.000)and the migration rate of cells wound healing was(56.37 ± 2.06)%,(25.00 ± 1.71)% sepa-rately(t=20.285,P=0.000).Conclusion Nifedipine may promote children's liver injury through regulating cell cycle related proteins.

Objective To validate the performance of six enzymatic glycated albumin reagents and evaluate their clinical application.Methods The performance of six enzymatic glycated albumin reagents(labled as A,B,C,D,E,F) from Beijing Jiuqiang Co, Beijing Lideman Co,Ningbomeikang Co, Beijing Haomai Co, Sichuan Maike Co and Asahi Kasei Co were assessed on Olympus AU5800 automatic biochemistry analyzer.According to the standard of CLSI,the precision,interference and linear correlation of these reagents were assessed.To assess the accuracy of GA% ,we used GA standard material whose value had been assigned using ID-LC/MS method provided by ReCCS.To do the method comparison and determine the consistency of assay, 50 fresh serum samples of T2DM outpatient and 80 fresh serum samples of apparently healthy people in Jan 2016 were tested using six kits.According to the EP28-A3C protocol, the reference range for GA%was validated in 122 apparently healthy individuals undertaking medical examination from January to February 2016 in PUMC.Results The precision,and the ability of anti-interference of the six reagents were good.The accuracy percentage deviation of six reagents was-19.3%-9.2%.The correlation coefficient of domestic reagents A to E and imported reagents F in the determination of GA% was 0.966-0.999, the average absolute bias was 7.0%-10.4%.The coincidence rate of A-E and F in determining abnormal GA% was between 88.5% and 96.9%.The coincidence rate was increased after switching to the reference range for preliminary clinical evaluation.Conclusion Six GA enzymatic kits used in automatic biochemical analyzer have high precision and strong anti-interference ability.Accuracy still needs to be improved.

Objective To investigate the relationship between maintaining concentration and loss of consciousness (LOC) concentration of propofol target controlled infusion (TCI) in patients undergoing heart valve replacement.Methods Thirty patients undergoing elective heart valve replacement were enrolled to receive propofol by ladder plasma TCI for anesthesia induction,8 males and 22 females.The initial plasma concentration (Cp) of propofol was set to 1.0 μg/ml,0.3 μg/ml Cp was increased every 1 min until LOC when the prediction effect-cite concentration (Ce) reached 0.5 μg/ml,then sufentanil 0.8-1.0 μg/kg and rocuronium 0.6-0.9 mg/kg were given for intubation.When BIS reached 50,Cp was decreased to the level of Ce.All the surgeries were performed under hypothermia CPB.MAP,HR,CVP,CO,SV,SVR,BIS,propofol Cp and Ce values were recorded at baseline (T0),LOC (T1),BIS reached 50 (T2),and other time points during operation (T3-T9).The correlation analysis between propofol Ce at LOC and perioperative variables were completed.Results In correlation analysis,there was a significant positive correlation between propofol Ce at LOC and baseline CO,SV (P<0.01),there was a significant negative correlation between propofol Ce at LOC and age (P<0.05),there was a significant positive correlation between propofol Ce at LOC and propofol Ce at T2-T9(P<0.01).Conclusion In patients undergoing valvular replacement,the Ce of propofol at maintenance are related to the concentration of propofol at LOC,which is helpful for adjusting the Ce of propofol at maintenance according to the Ce of propofol at LOC.