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 observe the pharmacokinetic effect of amoxicillin and clavulanate potassium tablets on amoxicillin in Chinese healthy subjects under fasting and high fat and high calorie diet.Methods 71 healthy subjects were given a single dose of amoxicillin potassium clavulanate tablets(0.375 g)on fasting or high fat diet,and venous blood samples were collected at different time points.The concentrations of amoxicillin in human plasma were determined by HPLC-MS/MS method,and the pharmacokinetic parameters were calculated by non-atrioventricular model using PhoenixWinNonlin 8.0 software.Results The main pharmacokinetic parameters of amoxicillin potassium clavulanate tablets after fasting and high fat diet were(5 105.00±1 444.00),(4 593.00±1 327.00)ng·mL-1,and postprandial-fasting ratio 89.40％,90％confidence interval(79.55％-100.19％);t1/2 were(1.52±0.16),(1.39±0.22)h;AUC0-t were(12 969.00±1 841.00),(11 577.00±1 663.00)ng·mL-1·h,and postdietary/fasting ratio 89.20％,90％confidence interval(83.92％-94.28％);AUC0-∞ were(13 024.00±1 846.00),(11 532.00±1 545.00)ng·mL-1·h,and postprandial-fasting ratio 88.60％,90％confidence interval(83.48％-93.50％).The median Tmax(range)were 1.63(0.75,3.00)and 2.50(0.75,6.00)h,respectively,and the Tmax of postprandial medication was delayed(P＜0.01).Conclusion Compared with fasting condition,amoxicillin Tmax was significantly delayed after high fat diet,while Cmax,AUC0-t and AUC0-∞ were not significantly changed,indicating that food could delay the absorption of amoxicillin,but did not affect the degree of absorption.

Objective To explore the effect and mechanism of salvianolic acid B on bronchial epithelial cell apoptosis induced by cigarette smoke extract.Methods Human bronchial epithelial cells 16HBE were divided into control group,model group(induced by cigarette smoke extract),experimental low-dose group(induced by cigarette smoke extract+11 μmol·L-1 salvianolic acid B treatment),and experimental medium-dose group(cigarette smoke extract Induction+22 μmol·L-1 salvianolic acid B treatment),experimental high-dose group(cigarette smoke extract induction+44 μmol·L-1 salvianolic acid B treatment),experimental high-dose+Compound C group(cigarette smoke extract induction+44 μmol·L-1 Salvianolic acid B+AMPK signaling pathway inhibitor Compound C treatment).Cell counting kit-8(CCK-8)assay was used to detect proliferation;Western blot was used to detect the phosphorylated adenosine monophosphate activates protein kinase(p-AMPK),CCAAT/enhancer-binding protein C/EBP(CHOP),activating transcription factor 4(ATF4)protein expression;PI single staining was used to detect cell cycle;Annexin V-FITC/PI double staining was used to detect apoptosis,and spectrophotometry was used to detect caspase-3 activity.Results The cell proliferation activity(OD value)in the bronchial epithelial cells of the control group,model group,experimental low-dose group,experimental medium-dose group,experimental high-dose group,and experimental high-dose+Compound C group were 0.86±0.07,0.38±0.03,0.45±0.03,0.54±0.04,0.68±0.03 and 0.42±0.04;the expression levels of p-AMPK/AMPK protein were 0.41±0.03,0.13±0.03,0.20±0.02,0.28±0.04,0.36±0.04 and 0.22±0.02;G0/G1 phase were(54.40±5.84)％,(82.93±4.50)％,(75.45±4.73)％,(67.41±2.70)％,(59.15±3.73)％and(69.80±6.59)％;apoptosis rate were(3.21±0.49)％,(24.90±3.35)％,(20.56±1.73)％,(13.55±1.68)％,(9.20±1.07)％and(18.04±1.79)％.Compared experimental low-dose group,experimental medium-dose group,experimental high-dose group with model group,the difference of above indicators were all statistically significant(all P＜0.05);compared the experimental high-dose+Compound C group with the experimental high-dose group,the difference of above indicators were all statistically significant(all P＜0.05).Conclusion Salvianolic acid B affects endoplasmic reticulum stress by activating AMPK signaling to reduce bronchial epithelial cell apoptosis induced by cigarette smoke extract.

Objective To explore the effect of forsythin on the Toll-like receptor 4(TLR4)/myeloid differentiation factor 88(MyD88)signaling pathway in mice infected with respiratory syncytial virus(RSV).Methods A respiratory infection model was established by intranasal inoculation of RSV.Mice infected with RSV were randomly divided into model group,experimental group,and positive control group,while control group received an equal volume of DMEM medium by nasal drops.The experimental group was administered forsythin(200 mg·kg-1·d-1)by gavage,the positive control group received ribavirin(27.6 mg·kg-1·d-1)by gavage,and both the control and model groups were given an equal volume of 0.9％NaCl,with continuous intervention for 14 days.Interleukin-4(IL-4),IL-13,and interferon-gamma(INF-γ)levels in bronchoalveolar lavage fluid(BALF)were detected by enzyme-linked immunosorbent assay;transcription levels of TLR4 and MyD88 mRNA in lung tissue were detected by real-time fluorescent quantitative polymerase chain reaction;and the positive expression of TLR4 and MyD88 proteins in lung tissue were detected by immunohistochemistry.Results The levels of IL-4 in BALF of the control group,model group,experimental group,and positive control group were(490.63±27.45),(1 382.37±41.28),(970.32±36.01),(738.41±30.27)pg·mL-1,respectively;IL-13 levels were(4.02±2.63),(18.33±5.62),(12.97±3.73),and(7.51±2.74)pg·mL-1,respectively;INF-γ levels were(54.42±2.68),(30.26±4.68),(42.80±3.36),and(45.01±3.82)pg·mL-1,respectively;the expression of TLR4 mRNA in lung tissue were 0.43±0.06,1.63±0.12,0.98±0.10,and 0.86±0.06,respectively;the expression of MyD88 mRNA in lung tissue were 0.56±0.09,1.78±0.15,1.02±0.07,and 0.75±0.06,respectively;the average optical density of TLR4 protein in lung tissue were 0.02±0.00,0.11±0.02,0.06±0.01,and 0.03±0.00,respectively;the average optical density of MyD88 protein were respectively 0.01±0.00,0.09±0.02,0.05±0.01,and 0.03±0.01,respectively.Compared the model group with the control group,and compared the experimental and positive control groups with the model group,above indicators showed statistically significant differences(all P＜0.05).Conclusion Forsythin can alleviate lung damage in mice infected with RSV,possibly by regulating the balance of immune cells and inhibiting the TLR4/MyD88 signaling pathway.

Objective To investigate the impact of anti-reflux modified duodenal exclusion surgery on glucose metabolism in rats with type 2 diabetes mellitus (T2DM), and to elucidate the role of the duodenum in maintaining glucose homeostasis. MethodsForty male Sprague-Dawley rats aged 5 weeks were fed a high-fat diet and induced with T2DM using low-dose streptozotocin. Thirty-six rats that met the T2DM model criteria were randomly divided into three groups: the simple duodenal exclusion surgery group (DE group), the anti-reflux modified duodenal exclusion group (MDE group), and the sham operation group (SO group), with 12 rats in each group. Gastroenterography was performed 4 weeks after surgery, and the body weight, fasting blood glucose levels, and serum glucagon-like peptide-1 (GLP-1) concentrations were measured before surgery and at 1, 2, 4, and 8 weeks post-surgery. Eight weeks post-surgery, the rats were euthanized, and a 1 cm segment of the biliopancreatic loop was collected from each group for pathological sectioning and HE staining to observe the intestinal mucosal villus length under an optical microscope. Results Gastroenterography showed that there was significant reflux of the contrast agent into the duodenal lumen in the DE group, while no reflux was observed in the MDE group. At one week post-surgery, the body weights of rats in all three groups significantly decreased compared to before surgery (P<0.05), and then the body weights of all groups increased over time, with no significant differences between the groups (P>0.05). Compared with the SO group, the fasting blood glucose levels in the MDE and DE groups significantly decreased at all time points post-surgery (P<0.05), while GLP-1 concentrations significantly increased (P<0.05). The fasting blood glucose levels in the MDE group were lower than those in the DE group at all time points post-surgery (P<0.05), but there were no significant differences in serum GLP-1 concentrations between the MDE and DE groups (P>0.05). Regarding intestinal mucosal morphology, the villus lengths of the biliopancreatic loops in the MDE group were significantly shorter than those in the DE and SO groups (P<0.05). Conclusion Anti-reflux modified duodenal exclusion surgery effectively improves glucose metabolism in T2DM rats by preventing the reflux of chyme into the diverted duodenum, thereby enhancing its hypoglycemic effect.

Objective: To explore the changes in coronal imbalance (CIB) in Lenke 5C adolescent idiopathic scoliosis (AIS) after posterior selective fusion surgery and determine their implications for surgical decision-making. Methods: One hundred twenty patients were categorized according to the preoperative coronal pattern (type A, coronal balance distance [CBD]<20 mm; type B, CBD≥20 mm and coronal C7 plumbline [C7PL] shifted to the concave side of the curve; type C, CBD≥20 mm and C7PL shifted to the convex side of the curve). CIB group (CIB+) was defined as having a CBD≥20 mm at the 2-year follow-up. Results: Compared to type A patients, the prevalence of postoperative CIB was higher in type C patients both immediately postoperative (22% vs. 38%, p<0.05) and at the final follow-up (5% vs. 29%, p<0.05), whereas type A patients showed a greater improvement in CBD (9 of 12 vs. 6 of 24, p<0.05) at the final follow-up. The majority of patients in all groups had recovered to type A at the final follow-up (96 of 120). The proximal Cobb-1 strategy reduced the incidence of postoperative CIB (1 of 38) at the 2-year follow-up, especially in preoperative type C patients. Multivariate logistic regression analysis revealed that type C and overcorrection of the thoracolumbar curve were risk factors for CIB at the 2-year follow-up (p=0.007 and p=0.026, respectively). Conclusion Patients with type C CIB in AIS exhibited unsatisfactory restoration, with 29% of them exhibiting CIB at the final follow-up. The selective fusion strategy of proximal Cobb-1 may reduce the risk of postoperative CIB especially when the preoperative coronal pattern is type C.

Objective:To analyze the effectiveness and interobserver agreement of the Parer five-tier, the National Institute of Child Health and Human Development (NICHD) three-tier, and the International Federation of Gynecology and Obstetrics (FIGO) three-tier electronic fetal monitoring (EFM) systems in identification of neonatal acidosis during labor.Methods:This retrospective study was conducted on full-term singleton cephalic deliveries with neonatal acidosis (umbilical artery blood gas pH≤7.1) and normal newborns (umbilical artery blood gas pH≥7.2) in the Nanjing Drum Tower Hospital, Nanjing University Medical School from January to December 2020. EFM tracings during the last 30-60 min before delivery were collected. Four obstetricians independently described the features of randomly sorted and coded EFM tracings. Another obstetrician categorized these tracings using the NICHD three-tier, FIGO three-tier, and Parer five-tier evaluation systems based on the features. All researchers were masked to the clinical characteristics and maternal and neonatal outcomes. The sensitivity and specificity for identifying neonatal acidosis, as well as the interobserver agreement, were analyzed for all three systems. Independent sample t-test, Chi-square (or Fisher's exact test) and Mann-Whitney U tests were used for statistical analysis. Inter-group comparisons of sensitivity and specificity between the three evaluation systems were assessed using McNemar's test. The Kappa statistic was used to analyze interobserver agreement. Results:This study included a total of 3 558 cases. After propensity score matching, there were 44 cases of neonatal acidosis and 78 control cases. There were no significant differences in parity, gestational weeks, modes of delivery, placental abruption, or analgesia rates between the two groups. The rates of instrumental vaginal delivery and neonatal intensive care unit (NICU) admission in the acidosis group were significantly higher than those in the control group [15.8% (7/44) vs. 2.6% (2/78), χ2=8.45, P=0.003; 31.8% (14/44) vs. 12.8% (10/78), χ2=8.45, P=0.004], while the umbilical artery blood pH and mean base excess were lower in the acidosis group than in the control group [7.04±0.07 vs. 7.30±0.05, t=4.98; (－12.40±3.32) vs. (－5.64±1.95) mmol/L, t=13.61; both P<0.001]. (2) Using the NICHD three-tier system, 95.5% (42/44) of the acidosis cases and 89.7% (70/78) of the control cases were classified as having category Ⅱ EFM tracings, indicating potential fetal acid-base imbalance; category Ⅲ EFM tracings were only observed in 4.5% (2/44) of the cases in the acidosis group. With the FIGO three-tier system, 81.8% (36/44) of the acidosis cases were categorized as having "pathological" tracings, and with the Parer five-tier system, 86.4% (38/44) of the acidosis cases were correctly classified into the "orange or red" risk zones that indicated acid-base imbalance. Among the control cases, there were 28.2% (22/78) with EFM tracings of "normal patterns" categorized by the FIGO three-tier system, and 41.0% (32/78) classified into the "green or blue" risk zones by the Parer five-tier system, which indicated good fetal conditions. None of the acidosis cases were misdiagnosed as being normal by the Parer five-tier system. (3) Compared with the NICHD three-tier system, both the FIGO three-tier and the Parer five-tier systems showed increased diagnostic sensitivity [4.5% (1.2%- 14.5%) vs. 81.8% (66.8%-89.4%) and 86.4% (71.8%-92.4%)], but decreased specificity [100.0% (95.3%- 100.0%) vs. 87.2% (78.0%-92.9%) and 84.6% (75.0%-91.0%)]. There was no statistically significant difference in the sensitivity or specificity between the FIGO three-tier and Parer five-tier systems for identifying neonatal acidosis ( P=0.727 and 0.791). (4) When reading the tracings of control cases, the total agreement rate for the NICHD three-tier system by different observers was as high as 94.2%, while the total agreement rates for the FIGO three-tier and Parer five-tier systems were 69.7% and 67.7%, respectively. In the interpretation of EFHR tracings for acidosis cases, the interobserver agreement for the Parer five-tier system was excellent [Kappa (95% CI): 0.87 (0.79-0.95)], while both the NICHD three-tier and FIGO three-tier systems showed good agreement [Kappa (95% CI): 0.77 (0.66-0.88) and 0.72 (0.60-0.84)]. Conclusions:The Parer five-tier and the FIGO three-tier systems have higher sensitivity in identifying neonatal acidosis than the NICHD three-tier system, and the Parer five-tier system achieves a higher negative predictive value and a greater agreement in the interpretation of pathological EFM patterns.

infC gene encodes the translation initiation factor IF3 in bovine Pasteuella multocida,but it whether or not regulation to the virulence and cross-protection in P.multocida is still not well understood.In this study,the infC gene mutant(△infC)derived from bovine P.multocida type A strain CQ2 was constructed using by homologous recombination method.Compared with wild strain,the △infC showed significant increasing in biofilm formation,but the capsule produc-tion,virulence and bacterial loading in organs were significant decreased,and the IL-1β secretion of mouse peritoneal macrophage increased.Along with the infC gene deletion,the expression of genes related to capsule synthesis and LPS synthesis and transport were significantly down-regulated,while that of genes related to biofilm synthesis and outer membrane protein were significantly up-regulated.The inactivated vaccines of wild type and mutant were prepared and mice were immu-nized twice then challenged with wild type strains,respectively.The immuno-protection rate of△in fC inactivated vaccine against bovine P.multocida type A,B and F were 100.0％,83.3％and 0.0％,respectively,and the immuno-protection rate that against rabbit type P.multocida was 33.3％.The results indicated that infC gene could affect the virulence of P.multocida by regula-ting the production of capsule and the expressions of virulence related factors,and the deletion of infC gene conferred a certain cross-protection property of strains.This study provided a certain foundation for the development of P.multocida vaccine.

Objective: To explore the changes in coronal imbalance (CIB) in Lenke 5C adolescent idiopathic scoliosis (AIS) after posterior selective fusion surgery and determine their implications for surgical decision-making. Methods: One hundred twenty patients were categorized according to the preoperative coronal pattern (type A, coronal balance distance [CBD]<20 mm; type B, CBD≥20 mm and coronal C7 plumbline [C7PL] shifted to the concave side of the curve; type C, CBD≥20 mm and C7PL shifted to the convex side of the curve). CIB group (CIB+) was defined as having a CBD≥20 mm at the 2-year follow-up. Results: Compared to type A patients, the prevalence of postoperative CIB was higher in type C patients both immediately postoperative (22% vs. 38%, p<0.05) and at the final follow-up (5% vs. 29%, p<0.05), whereas type A patients showed a greater improvement in CBD (9 of 12 vs. 6 of 24, p<0.05) at the final follow-up. The majority of patients in all groups had recovered to type A at the final follow-up (96 of 120). The proximal Cobb-1 strategy reduced the incidence of postoperative CIB (1 of 38) at the 2-year follow-up, especially in preoperative type C patients. Multivariate logistic regression analysis revealed that type C and overcorrection of the thoracolumbar curve were risk factors for CIB at the 2-year follow-up (p=0.007 and p=0.026, respectively). Conclusion Patients with type C CIB in AIS exhibited unsatisfactory restoration, with 29% of them exhibiting CIB at the final follow-up. The selective fusion strategy of proximal Cobb-1 may reduce the risk of postoperative CIB especially when the preoperative coronal pattern is type C.

Objective: To explore the changes in coronal imbalance (CIB) in Lenke 5C adolescent idiopathic scoliosis (AIS) after posterior selective fusion surgery and determine their implications for surgical decision-making. Methods: One hundred twenty patients were categorized according to the preoperative coronal pattern (type A, coronal balance distance [CBD]<20 mm; type B, CBD≥20 mm and coronal C7 plumbline [C7PL] shifted to the concave side of the curve; type C, CBD≥20 mm and C7PL shifted to the convex side of the curve). CIB group (CIB+) was defined as having a CBD≥20 mm at the 2-year follow-up. Results: Compared to type A patients, the prevalence of postoperative CIB was higher in type C patients both immediately postoperative (22% vs. 38%, p<0.05) and at the final follow-up (5% vs. 29%, p<0.05), whereas type A patients showed a greater improvement in CBD (9 of 12 vs. 6 of 24, p<0.05) at the final follow-up. The majority of patients in all groups had recovered to type A at the final follow-up (96 of 120). The proximal Cobb-1 strategy reduced the incidence of postoperative CIB (1 of 38) at the 2-year follow-up, especially in preoperative type C patients. Multivariate logistic regression analysis revealed that type C and overcorrection of the thoracolumbar curve were risk factors for CIB at the 2-year follow-up (p=0.007 and p=0.026, respectively). Conclusion Patients with type C CIB in AIS exhibited unsatisfactory restoration, with 29% of them exhibiting CIB at the final follow-up. The selective fusion strategy of proximal Cobb-1 may reduce the risk of postoperative CIB especially when the preoperative coronal pattern is type C.