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 compare the clinical features, clinical efficacy, and prognosis of patients with double-hit and non-double-hit high-risk multiple myeloma (MM) and explored the clinical significance of high-risk cell karyotype in MM development.Methods:The clinical data of 73 high-risk MM patients admitted to the Department of Hematology of Fujian Provincial Hospital from January 2011 to February 2019 were retrospectively analyzed. Interphase fluorescence in situ hybridization was used to detect their karyotypes. Based on mSMART 3.0 risk stratification, we divided the patients into a double-hit group (28 cases) and a non-double-hit group (45 cases).Results:Fifteen patients in the double-hit group and 26 in the non-double-hit group received bortezomib-based chemotherapy. The median progression-free survival (PFS) in the double-hit and the non-double-hit groups was 8.0 months and 22.0 months, and the median overall survival (OS) was 10.0 months and not reached, respectively. Ten patients in the double-hit group and 12 in the non-double-hit group received bortezomib combined with lenalidomide (RVD) chemotherapy. The median PFS in the double-hit group and the non-double-hit group was 12.0 months and 24.0 months, and the median OS was 14.0 months and not reached, correspondingly. Both the PFS and OS of the double-hit group were significantly shorter than those of the non-double-hit group ( P<0.05). Univariate analysis results indicated that cytogenetic abnormalities, revised-international staging system (R-ISS), β2 microglobulin, and calcium had significant effects on PFS in high-risk MM patients ( P<0.05). The cytogenetic abnormalities, R-ISS, and β2 microglobulin were associated with OS in high-risk MM patients ( P=0.001). Multivariate Cox regression analysis showed that the cytogenetic grouping was an independent prognostic factor for OS and PFS in high-risk MM patients. The risk of disease progression was 3.160 times (95% CI: 1.364-7.318) and the risk of death was 2.966 times higher (95% CI: 1.205-7.306) in the double-hit group than those in the non-double-hit group. Calcium was an independent risk factor for PFS in the high-risk MM patients. Notably, the risk of disease progression in patients with calcium levels≥ 2.75 mmol/L was 2.667 times higher than that in patients with calcium<2.75 mmol/L (95% CI: 1.209-5.883). Conclusions:Double-hit patients are a highly specific group with worse high-risk MM prognosis. In such patients, the relapse is more common, the disease progression is faster, and the survival time is shorter than those in the non-double-hit patients.

Objective:To analyze the clinical features and efficacy of enzyme replacement therapy in 4 children with Fabry disease.Methods:A retrospective analysis of the clinical manifestations, laboratory findings, genetic variations and treatment were conducted in 4 children with Fabry disease in Children′s Hospital of Zhejiang University School of Medicine from January 2014 to July 2020.Results:All four children (2 males, 2 females) with onset age of 12.4 (6.0-16.8) years were diagnosed based on clinical features, α-Gal A enzyme activity, genetic analysis and family history. The clinical manifestations varied in 4 children. All patients had left ventricular hypertrophy and abnormal urinalysis results, 1 case of neuropathic pain, 2 cases of hypohidrosis, 1 case of insipidus, but no angiokeratomas or hearing abnormalities were found. Three missense mutations of GLA gene were identified: c.424T>C (p.C142R), C.335G>A (p.R112H) and c.644A>G (p.N215S). The first two gene mutations were classical phenotypes, and the last one had also been reported in a classic case. In Case 1, no severe adverse events were reported in the first two months of agalsidase beta treatment. The dosage was 1 mg/kg once every 2 weeks. Symptoms of pain intensity and hypohidrosis were improved. Transiently elevated proteinuria was observed but it returned to normal after a week without any treatment.Conclusions:Clinical manifestations of Fabry disease varied in childhood. Multidisciplinary collaboration is required for its early diagnosis and treatment. Severe adverse events are rare in children with short-term therapy of agalsidase beta.

Objective:To compare the clinical features, clinical efficacy, and prognosis of patients with double-hit and non-double-hit high-risk multiple myeloma (MM) and explored the clinical significance of high-risk cell karyotype in MM development.Methods:The clinical data of 73 high-risk MM patients admitted to the Department of Hematology of Fujian Provincial Hospital from January 2011 to February 2019 were retrospectively analyzed. Interphase fluorescence in situ hybridization was used to detect their karyotypes. Based on mSMART 3.0 risk stratification, we divided the patients into a double-hit group (28 cases) and a non-double-hit group (45 cases).Results:Fifteen patients in the double-hit group and 26 in the non-double-hit group received bortezomib-based chemotherapy. The median progression-free survival (PFS) in the double-hit and the non-double-hit groups was 8.0 months and 22.0 months, and the median overall survival (OS) was 10.0 months and not reached, respectively. Ten patients in the double-hit group and 12 in the non-double-hit group received bortezomib combined with lenalidomide (RVD) chemotherapy. The median PFS in the double-hit group and the non-double-hit group was 12.0 months and 24.0 months, and the median OS was 14.0 months and not reached, correspondingly. Both the PFS and OS of the double-hit group were significantly shorter than those of the non-double-hit group ( P<0.05). Univariate analysis results indicated that cytogenetic abnormalities, revised-international staging system (R-ISS), β2 microglobulin, and calcium had significant effects on PFS in high-risk MM patients ( P<0.05). The cytogenetic abnormalities, R-ISS, and β2 microglobulin were associated with OS in high-risk MM patients ( P=0.001). Multivariate Cox regression analysis showed that the cytogenetic grouping was an independent prognostic factor for OS and PFS in high-risk MM patients. The risk of disease progression was 3.160 times (95% CI: 1.364-7.318) and the risk of death was 2.966 times higher (95% CI: 1.205-7.306) in the double-hit group than those in the non-double-hit group. Calcium was an independent risk factor for PFS in the high-risk MM patients. Notably, the risk of disease progression in patients with calcium levels≥ 2.75 mmol/L was 2.667 times higher than that in patients with calcium<2.75 mmol/L (95% CI: 1.209-5.883). Conclusions:Double-hit patients are a highly specific group with worse high-risk MM prognosis. In such patients, the relapse is more common, the disease progression is faster, and the survival time is shorter than those in the non-double-hit patients.

Objective To evaluate value of narrow band imaging(NBI) endoscopy for children with abdominal Henoch?Schonlein purpura ( HSP ) . Methods A total of 46 patients with abdominal HSP were enrolled into the observation group(NBI intervention) from November 2010 to February 2016.Diagnostic rates of white light and NBI endoscopy in abdominal HSP patients, IgA positive rates of targeted biopsies and severe complications were retrospectively analyzed. A total of 25 abdominal HSP patients with no NBI intervention admitted from 2007 to 2009 were randomly enrolled into control group. Data of the control group were compared with those of observation group. Results In observation group, the diagnostic rate under NBI was significantly higher than that under white?light endoscopy[91. 3%(42/46)VS 67. 4%(31/46),χ2=8. 02,P<0. 05]. IgA positive rates of targeted biopsies under NBI was significantly higher than that under white?light endoscopy [ 95. 7%( 88/92 ) VS 69. 6%( 64/92 ) ,χ2 = 21. 79, P<0. 05 ] . Three patients developed such serious complications as digestive hemorrhage as predicted. Compared with control group, abdominal pain and blood stool relief time (10. 96±5. 32 d VS 19. 68±4. 29 d,t=7. 50,P<0. 01), fasting time(10. 37±5. 42 d VS 8. 80± 3. 71 d,t=7. 73,P<0. 01), hospital stay (18. 80±7. 11 d VS 23. 12±4. 36 d, t=3. 16,P<0. 01), time of stool occult blood negative ( 11. 41 ± 6. 30 d VS 19. 12 ± 4. 09 d, t=6. 22, P<0. 01 ) in observation group were significantly shortened. Conclusion NBI endoscopy is valuable for improving the diagnostic accuracy and biopsy accuracy and complication prediction of abdominal Henoch?Schonlein purpura in children.

Objective To compare the application values of different blood transfusion modes in placenta previa cesarean section.Methods The clinical data in 82 pregnant women with placenta previa undergoing cesarean section in our hospital from February 2013 to January 2016 were collected.The patients were divided into the autologous group (autologous blood stored blood transfusion,n=42) and allogeneic group (allogeneic blood transfusion,n=40) according to different blood transfusion modes.The changes of blood routine indexes such as hemoglobin (Hb),platelet count (PLT),hematocrit (Hct),white blood cell count (WBC) and red blood cell count (RBC) in the two groups before and after operation were recorded.Postpartum blood lossb amounts,autologous and allogeneic blood transfusion volume were compared between the two groups.The pregnant outcomes were observed,and the incidence rates of blood transfusion complications were statistically analyzed.Results (1)The accumulative blood loss volume and allogeneic blood transfusion volume in the autologous group were significantly lower than those in the allogeneic group (P<0.05);(2) PLT and WBC after operation in the two groups were significantly increased,while RBC,Hb and Hct were decreased.The levels of postoperative Hb,PLT and Hct in the autologous group were higher than those in the allogeneic group (P<0.05);(3) the neonatal Apgar scores at 1,5 min after birth,and umbilical artery blood pH value showed no statistically significant difference between the two groups (P>0.05);(4)The total incidence rate of complications in the autologous group was significantly lower than that in the allograft group (P<0.05).Conclusion Adopting autologous blood stored blood transfusion scheme during cesarean section in women with placenta previa has no negative effect on maternal and neonatal outcomes,meanwhile which can reduce the incidence of transfusion complications,is safe and feasible.

Objective To investigate the complications and management of nitinol self-expandable metal stent (cSEMS) in treatment of refractory esophageal strictures in children. Methods The clinical data were reviewed for 9 pediatric patients with refractory benign esophageal disorders from May 2009 to December 2016, specially designed cSEMS were applied to them, data about effects and complications were collected during regular follow-ups. Results Successful cSEMS placement was performed in 9 children, the symptom of dysphagia was obviously alleviated after implantation, all patients underwent vomiting and chest pain 1~7 days after operation; 1 case could not put up with the pain, so the stent had to be removed in 36 hours after implantation; 2 cases developed a recurrent stricture within 3 months after stent removal, growth of mild granulation tissue was found in 1 case; In the case with esophageal fistulas, migration and poor adherence to the esophagus was occurred in 3 days after implantation, then a new designed cSEMS with bigger proximal tip was planted in the same place 1 week later, 2 months after stent removed, fistula was healed. Conclusion Placement of cSEMS is safe and effective in treating pediatric patients with refractory esophageal stricture. However, complications associated with stent placement should not be ignored, individually designed stent and timely management of the complications are quite important in order to enhance clinical efficacy.

Objective To evaluate the value of endoscopy with narrow-band imaging(NBI)for tar-geting biopsy of lesions in children with allergic colitis. Methods The colorectal lesions under the model of traditional white light imaging and NBI in children with allergic colitis were observed and recorded. The re-sults of biopsy under white light imaging and NBI were compared with postoperative pathological diagnosis. Results A total of 60 patients completed the study. Under NBI,47(78. 3%)rectal lesions,52(86. 7%) sigmoid colon lesions and 19(31. 7%)descending colon lesions were found. There were 21(35. 0%),25 (41. 7%),and 8(13. 3%)lesions in the transverse colon,ascending colon and iloececus,respectively. For the lesions in transverse colon and ascending colon,the detection rate under NBI was significantly higher than that under white light endoscopy[35. 0% VS 16. 7%(10/ 60),P<0. 05;41. 7% VS 18. 3%(11/ 60),P<0. 05]. No significant difference was found between two methods for the sigmoid and rectal lesion[86. 7% VS 83. 3%(50/ 60),P>0. 05;78. 3% VS 76. 7%(46/ 60),P>0. 05].There was no significant difference in ileo-cecus and descending colon[13. 3% VS 5. 0%(3/ 60),P>0. 05;31. 7% VS 28. 3%(17/ 60),P>0. 05]. The targeted biopsy results by NBI indicated that the ratio of EOS≥6/ HP was significantly higher than the biopsy results by white light imaging. Conclusion NBI is easy to operate and can discern the microcosmic structure which cannot be observed by the traditional endoscopy. It can provide higher diagnostic accuracy of allergic colitis. After suspicious lesions were found under conventional endoscopic check,NBI could be used to guide the targeted biopsy.

AIM:To investigate whether Notch1 changes stemness and chemotherapeutic sensitivity in human glioma U251 cells.METHODS: The lentiviral vectors, which expressed Notch1-shRNA or Notch1 intracellular domain ( NICD) , were transfected into U251 cells .Western blot and immunofluorescence staining were applied to monitor the va-lidity of the cells, down-regulation of Notch1 expression or over-expression of NICD.The proportion of CD133 +cells was analyzed by flow cytometry.The expression of nestin and GFAP was identified by immunofluorescence staining.The forma-tion rate of tumor cell spheres and the implanted tumor growth in SCID mice were observed.MTT assay was performed to e-valuate the chemotherapeutic sensitivity to VM-26 and BCNU of the cells with different treatments.RESULTS:Stemness was significantly enhanced in the cells over-expressing NICD.For example, the proportion of CD133 +cells was increased, the expression of nestin was up-regulated, the expression of GFAP was down-regulated, and the formation rate of tumor cell spheres and implanted tumor growth were increased.The chemotherapeutic sensitivity to VM-26 and BCNU of the cells was decreased.In the cells with Notch1 gene down-regulation by RNAi, the stemness was inhibited and chemotherapeutic sensi-tivity was increased.CONCLUSION:Notch1, which leads to the change of stemness and chemotherapeutic sensitivity in human glioma U251 cells, is likely to be a potential molecular target for treatment of glioma.

Objective To assess the interference by calcium dobesilatein 7 peroxidase-baseduric acid assays and to determine its clinical significance.Methods In the in vitro experiments, uric acid in pooled serum with final concentrations of calcium dobesilate additions (0, 2, 4, 8, 16, 32, and 64μg/ml) were measured by 7 peroxidase-based assays.Percent Bias (%) was calculated relative to the drug-free specimen.In the in vivo experiments, changes in serum uric acid and calcium dobesilate concentrations were observed before and after calcium dobesilate administration ( baseline, 0 h, 1 h, 2 h, 3 h, 4 h, 6 h ) involunteers.The interference in different assays was assessed compared with LC-IDMS/MS method. Calcium dobesilate levels in 40 specimens from those taking calcium dobesilate were measured by HPLC method.Of the 40 specimens, 10 were selected to analyse the levels of uric acid by both peroxidase and UV measurement method to assess the impact in clinical status.Results In the in vitro study, concentrations of uric acid measured by 7 peroxidase-based assays were reduced by -6.3%to -21.2%compared with drug-free serum, when theconcentration of calcium dobesilate was16μg/ml.In the in vivo study, comparedto UA levels at 0 h, the biasesof serum uric acid determined by peroxidase method after calcium dobesilate administration(1 h, 2 h, 3 h, 4 h, 6 h) were of -3.33%, -6.79%, -7.49%, -6.07%, -4.09%, respectively.The observed uric acid concentrations for 8 participants measured by enzymatic assays were inhibited by -3.75% to -6.89% at 0 hour and by -16.9% to-22.22% at 2 hours relative to the concentrations measured by the LC-IDMS/MS method. Conclusions Calcium dobesilate produced a clinically significant negative interference with uric acid in all peroxidase-based uric acid assays,which may result in false evaluation of uric acid level in clinical status.Significant differences in the degree of interference were observed among the assays.