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 develop a new approach for the preparation of 0.7~0.8 hematocrit concentrated washed red blood cells(RBCs) for fetal anemia in utero transfusion and apply it in clinical. 【Methods】 The erythrocyte suspension and frozen stored erythrocytes within expiry date in Guangzhou Blood Center from March 2020 to February 2021 were taken to prepare concentrated washed RBCs. According to the derivation formula, corresponding weight of RBC preservation solution was added to obtain 0.7~0.8 hematocrit concentrated washed RBCs. Routine blood test data were statistically analyzed by single-sample t test, and P<0.05 was considered statistically significant. Qualified Rh-negative/ O-type 0.7~0.8 hematocrit concentrated RBCs within expiry date were used in clinical intrauterine blood transfusion. 【Results】 The hematocrit of concentrated washing RBCs prepared by the new approach could reach 0.7~0.8. The RBCs count (8.389 ±0.808)×1 0¹²/ L and hemoglobin content(233.730±15.498)g/L were higher while the erythrocyte count (0.732±0.469)×10⁹ /L and platelets count(26.000±26.276)×10⁹/L were lower than the normal values of adults. The mean erythrocyte volume(fL), hemoglobin content(pg) and concentration(g/L )were 88.123±6.359, 30.004±2.809 and 339.980±11.865, respectively, which were normal values of adults. Fetal anemia was significantly improved and the prognosis was good after intrauterine blood transfusion. 【Conclusion】 The 0.7~0.8 hematocrit concentrated washed RBCs prepared by the new approach is consistent with the special blood requirements during fetal anaemia transfusion, meets the clinical treatment standards, and can be applied in clinical.

Objective:To analyze the application of sensory stimulation based on the whole process accompanying of relatives in regain of consciousness in coma patients after aneurysm embolization.Methods:Comatose patients after aneurysm embolization in Department of Cerebrovascular Disease, Suining Central Hospital of Sichuan Province were selected as the research objects. 59 patients from June to December in 2019 were included in the control group and received routine sensory stimulation. 60 patients from January to August 2020 were included in the study group, and the sensory stimulation based on the whole process accompanying of relatives was implemented. Glasgow Coma Scale (GCS) , Chinese version of Coma Recovery Scale-Revised (CRS-R) and Mini-Mental State Examination (MMSE) were applied to compare the coma status, recovery status and cognitive function of the two groups before and after the intervention. The complications of the two groups were compared as well.Results:After 3 months of intervention, the scores of GCS, CRS-R and MMSE of the study group were all higher than those of the control group, with statistically significant differences ( P<0.05) . Within 3 months after operation, the total incidence of complications in the study group was lower than that in the control group, and the difference was statistically significant ( P<0.05) . Conclusions:Sensory stimulation based on the whole process accompanying of relatives is helpful to reduce the degree of coma, promote the recovery of patients, and reduce the incidence of complications.

Objective: By comparing different repairing effects of different methods on articular cartilage defects in rabbit model, the ability of a new bioactive glass scaffold to repair cartilage defect was studied. Methods: We prepared the PSC/CS bone cement by mixing the bioactive glass (BG) powder composed of 10.8%P₂O₅-54.2%SiO₂-35%CaO (PSC), chitosan solution (CS) and calcium sulfate hemihydrate (CSH), then the bone cement was sized by a mold to form cylindrical scaffold. In vivo experiment, 18 male rabbits were divided into three groups randomly, including blank group, BG group and TGF-β1 group, which was added TGF-β1 into BG scaffold. Both knees of each rabbit were made cartilage defect for the same group, and no intervention was applied in B group, then implanted scaffolds into defects in both experimental groups. At 6 and 12 weeks after surgery, observed the macroscopic growth, histologic staining and collagen II immunohistochemistry (IHC), and the International Cartilage Repair Society (ICRS) and Wakitani score were used to analyze the experimental results quantitatively. Results: At 6 weeks after surgery, there was no obvious difference between blank group and BG groups, but the macroscopic result of TGF-β1 group was better than the other two groups and its ICRS score 4.67±0.52 points was statistically higher than BG group 2.83±0.75 points (t=-2.817, P=0.015). As to the comparison of histologic staining and Wakitani socre among three groups, no statistical difference was observed (blank group=13.67±0.52, BG group=13.83±0.41, TGF-β1 group=13.33±1.03). At 12 weeks after surgery, there was still no obvious difference between blank and BG groups, while the results of macroscopic observation and ICRS score in TGF-β1 group were significantly higher than them 9.01±0.63 points (blank group vs TGF-β1 group: t=-2.289, P=0.022; BG group vs TGF-β1 group: t=-2.326, P=0.020). More importantly, much deeper positive staining were observed in TGF-β1 group, and the Wakitani score was higher than the other two groups (blank group=9.83±1.33, BG group=9.51±1.05, TGF-β1 group=6.50±1.38, blank group vs TGF-β1 group: t=-2.771, P=0.007; BG group vs TGF-β1 group: t=-2.756, P=0.006). By comparing the degree of histologic staining and Col II expression with normal cartilage, the regenerated tissue in TGF-β1 group was similar. Conclusion Single PSC/CS scaffold doesn’t possess excellent ability to repair cartilage defect. When TGF-β1 was added into PSC/CS bioactive glass, the scaffold was able to promote cartilage defect repair, and the regenerated tissue was similar to normal cartilage.

Objective To translate the Water Orientation Test of Alyn 2 (WOTA2) into Chinese, and to study its reliability and validity for patients with spinal cord injury. Methods After authorizing, the English version WOTA2 scale and its training package were translated into Chinese. From January to September, 2018, 137 patients with spinal cord injury were evaluated with the Chinese version WOTA2 by two evaluators independently, and evaluated again three days later by one of the evaluators. The Cronbach's α, Spearman-Brown coefficient, intra-group correlation coefficient (ICC) and Kappa coefficient were calculated to evaluate the reliability. Content Validity Index (CVI) of items (I-CVI) and scale (S-CVI) were used to evaluate the content validity, and factors analysis was used to evaluate the structure validity. Results The Cronbach's α was 0.947 in all items of the scale, 0.890 in mental adjustment items and 0.954 in aquatic skills items. Pearson correlation coefficient of the half scales separated by odd and even items was 0.948, and the Spearman-Brown coefficient was 0.973. The ICC of test-retest was 0.965 in total score, 0.965 in centesimal system total score, 0.847 in mental adjustment score and 0.970 in aquatic skills score. The ICC of inter-testers was 0.964 in total score, 0.965 in centesimal system total score, 0.847 in mental adjustment score and 0.970 in aquatic skills score. The Kappa coefficient was 0.528 to 0.927 in test-retest and 0.528 to 0.927 in inter-testers. The I-CVI was 0.8 to 1.0 and S-CVI was 0.63. Factors analysis extracted four factors, which met the theory, and contributed 67% of cumulative variance. Conclusion The Chinese version WOTA2 is good in reliability and validity for patients with spinal cord injury, and can be used in the clinical practice of aquatic therapeutic exercise in China.

Objective To campare the diagnostic value of contrast-enhanced ultrasound(CEUS) and International Ovarian Tumor Analysis (IOTA) simple rules in the differential diagnosis of adnexal tumors . Methods For the 101 suspicious malignant adnexal tumors discovered by conventional ultrasound ,IOTA simple rules and CEUS were performed before operation . Tumors were divided into benign ,uncertain and malignant according to IOTA simple rules . Tumors were scored by CEUS according to the membrane integrity ,coating thickness uniformity ,separation enhancement ,enhance strength ( no enhancement ,low enhancement ,equal or high enhancement) ,and enhanced performance( uniform and non-uniform) . The ROC curve was drawn with the pathological results as the gold standard ,and the best boundary value was obtained . Finally ,the diagnostic efficiency of the diagnostic methods were analyzed . Results ①According to the ROC curve analysis ,the area under the curve of IOTA simple rules was 0 .757( P ＝0 .000) . When the uncertainty type was classified as benign ,the random index was 0 .41 ,the sensitivity was 61 .54% ,and the specificity was 79 .59% . When the uncertainty type was classified as malignant ,the random index was 0 .31 ,the sensitivity was 96 .15% ,and the specificity was 34 .69% . When the uncertain types were excluded the Youden index was 0 .77 ,the sensitivity was 76 .19% ,and the specificity was 89 .47% . ②The differences in the membrane integrity , coating thickness uniformity , separation enhancement , enhance strength , enhanced performance between benign and malignant tumors were statistically significant( P ＜ 0 .01) . ③According to the ROC curve analysis ,the area under the curve of CEUS was 0 .914( P ＝ 0 .000) ,cut-off value was 3 score . The Youden index ,sensitivity and specificity of evaluating benign and malignant adnexal tumors were 0 .80 ,78 .85% and 91 .84% respectively . ④ The tumors evaluated as benign by IOTA simple rules was classified as benign . The tumors evaluated as malignant by IOTA simple rules were classified as malignant . The tumors evaluated as uncertain by IOTA simple rules and scored less than or equal to 3 points according to CEUS were classified as benign . The tumors evaluated as uncertain by IOTA simple rules and scored more than 3 points according to CEUS were classified as malignant . According to the ROC curve analysis ,the area under the curve of CEUS combined with IOTA simple rules was 0 .831 ( P ＝0 .000) . The Youden index ,sensitivity and specificity in evaluating benign and malignant adnexal tumors were respectively 0 .66 ,86 .54% and 79 .59% respectively . Conclusions CEUS has a higher diagnostic efficiency than IOTA Simple Rules and the two combined in the diagnosis of adnexal tumors .

Fluorescence spectroscopy was used to investigate the efficiency of coupling peptides to gold nanoparticles via 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride-N-hydroxysuccinimide (EDC-NHS).The experiment conditions including buffer solution, pH value and concentrations of buffer solution, concentrations of NHS and EDC, concentration ratios of NHS to EDC, and coupling reaction time on the coupling efficiency were investigated and optimized.The experimental results indicated that the optimized experimental conditions were as follows: 25 mmol/L HEPES buffer solution, pH 7.0;2∶1 of concentration ratio of NHS to EDC, 0.4 mol/L NHS, 0.2 mol/L EDC, and coupling reaction time of 24 h.This study may provide references for the relative research involving coupling peptide or protein with gold nanoparticles

Background It has not been reported that if the visual cortex M receptor changed during the development of myopia and how it changed if given acupuncture treatment.Objective The aim of this study was to observe the effect of electroacupuncture stimulation on the expression of acetylcholine receptors M1 (AchRM1) in visual cortex of guinea with lens-induced myopia (LIM).Methods Forty-eight three-week-old healthy guinea pigs were randomized into the normal control group,the LIM model group and the LIM electroacupuncture group.The right eyes of the guinea pigs were selected as the experimental eyes.LIM was created by monocularly wearing of-10 D lens for 4 weeks in the right eyes in the LIM model group and LIM electroacupuncture group,and then the acupuncture at the temple and hegu point was performed for 30 minutes per day for consequent 4 weeks,in the LIM electroacupuncture group.The fellow eyes of the guinea pigs were used as the self-control eyes.The refractive power and axial length were examined with retinoscopy and A-type sonography before and 4 weeks after modeling,respectively.The animals were sacrificed by excessive anesthesia at the fourth week after acupuncture and visual vertex tissue was obtained.The expression of M1 receptor mRNA in visual vertex was detected by fluorescence quantitative PCR,and the content of M1 receptor protein in visual vertex was assyed by ELISA.The study protocal was approved by Animal Ethic Committee of Shandong University of Traditional Chinese Medicine,and the use and care complied with Statement of the Association for Research in Vision and Ophthalmology.Results At the fourth week after modeling,the mean diopters were (-3.24±0.28) D and (-3.30±0.45) D in the LIM model group and the LIM eleetroacupuncture group,which were significantly higher than (0.83 ±0.86)D in the normal control group (both at P=0.000),and there was no significant difference in the diopter between the LIM model group and the LIM electroacupuncture group (t =0.200,P =0.659).The mean axial lengths were (8.67 ±0.14) mm and (8.60±0.06) mm in the LIM model group and the LIM electroacupuncture group,which were considerably increased in comparison with (8.33±0.08)mm in the normal control group (both at P＜0.05).The relative expression levels of AchRM1 mRNA in visual cortex were 0.79±0.18,1.36±0.23 and 1.13±0.13 in the normal control group,LIM model group and LIM electroacupuncture group,and the relative expression level of AchRM1 mRNA in the LIM electroacupuncture group was significantly higher than that of the normal control group and lower than that of the LIM model group (both at P＜0.05).In addition,the contents of AchRM1 receptor protein in the visual cortex were 248.00±33.31,455.17±42.40 and 396.17±47.57 in the normal control group,LIM model group and the LIM electroacupuncture group,with a similar pattern among the groups (both at P＜0.05).Conclusions A electroacupuncture stimmulation do not affect the myopic diopter and axial length in LIM model.The AchRM1 and AchRM1 receptor in the visual cortex up-regulate in LIM eyes,infering that electroacupuncture stimmulation can improve vision by decreasing the level of AchRM1 receptor in visual cortex in LIM eyes in guinea pigs.

Objective To describe the changing demographic profile over time of pregnant women diagnosed with HIV infection, and trends in mode of delivery and pregnancy outcome. Methods An observational investigation was conducted based on a population of 1 128 HIV-positive pregnant women at eight sites in China in provinces with high prevalence of HIV among pregnant women and children between 2007 and 2013. The study area included Shangcai and Weishi counties in Henan province; Ruili and Longchuan counties and Linxiang prefecture in Yunnan province;Yining County in Xinjiang;and Lingshan county and Babu district in Guangxi province. Main outcome measures were maternal characteristics, mode of delivery, and related trends over time by Cochram-Mantel-Haenszel(CMH) χ2-test. Results A total hospital delivery rate of 96.4%was reported. For each year during the period 2007-2013, hospital delivery rates were 95.1%(253/266), 94.8%(188/198), 93.7%(164/175), 98.8%(159/161), 98.6%(141/143), 97.4%(114/117), and 100.0%(68/68), respectively (χ2CMH=7.68, P=0.006). The neonatal survival rate increased by 0.7% incrementally from 2007 to 2013, with 98.5% (262/266), 99.0% (196/198), 99.4% (174/175), 99.4%(160/161), 100.0% (143/143), 100.0% (117/117), and 100.0% (68/68), respectively (χ2CMH=4.58, P=0.030). The proportions of vaginal deliveries in those years were 62.2% (166/266), 54.8% (109/198), 59.9% (105/175), 61.7% (100/161), 66.7% (96/143), 58.8% (69/117), and 66.7% (46/68), respectively (χ2CMH=1.19, P=0.276). The average rate for midwives skilled in procedures such as artificial rupture of membranes and episiotomy was 37.8% (261/691), with proportions decreasing during the study period at 39.2% (65/166), 47.4% (52/109), 35.9% (38/105), 46.0% (46/100), 25.5% (25/96), 31.2% (22/69), and 28.3% (13/46), respectively (χ2CMH=6.76, P=0.009). Conclusion The annual rate of reported hospital delivery increased between 2007 and 2013, with a declining rate of skilled midwifery over the same period. Survival of neonates born to HIV-infected mothers in China has increased to nearly equivalent levels as those of developed countries.

Objective To describe the changing demographic profile over time of pregnant women diagnosed with HIV infection, and trends in mode of delivery and pregnancy outcome. Methods An observational investigation was conducted based on a population of 1 128 HIV-positive pregnant women at eight sites in China in provinces with high prevalence of HIV among pregnant women and children between 2007 and 2013. The study area included Shangcai and Weishi counties in Henan province; Ruili and Longchuan counties and Linxiang prefecture in Yunnan province;Yining County in Xinjiang;and Lingshan county and Babu district in Guangxi province. Main outcome measures were maternal characteristics, mode of delivery, and related trends over time by Cochram-Mantel-Haenszel(CMH) χ2-test. Results A total hospital delivery rate of 96.4%was reported. For each year during the period 2007-2013, hospital delivery rates were 95.1%(253/266), 94.8%(188/198), 93.7%(164/175), 98.8%(159/161), 98.6%(141/143), 97.4%(114/117), and 100.0%(68/68), respectively (χ2CMH=7.68, P=0.006). The neonatal survival rate increased by 0.7% incrementally from 2007 to 2013, with 98.5% (262/266), 99.0% (196/198), 99.4% (174/175), 99.4%(160/161), 100.0% (143/143), 100.0% (117/117), and 100.0% (68/68), respectively (χ2CMH=4.58, P=0.030). The proportions of vaginal deliveries in those years were 62.2% (166/266), 54.8% (109/198), 59.9% (105/175), 61.7% (100/161), 66.7% (96/143), 58.8% (69/117), and 66.7% (46/68), respectively (χ2CMH=1.19, P=0.276). The average rate for midwives skilled in procedures such as artificial rupture of membranes and episiotomy was 37.8% (261/691), with proportions decreasing during the study period at 39.2% (65/166), 47.4% (52/109), 35.9% (38/105), 46.0% (46/100), 25.5% (25/96), 31.2% (22/69), and 28.3% (13/46), respectively (χ2CMH=6.76, P=0.009). Conclusion The annual rate of reported hospital delivery increased between 2007 and 2013, with a declining rate of skilled midwifery over the same period. Survival of neonates born to HIV-infected mothers in China has increased to nearly equivalent levels as those of developed countries.