Objective:To compare the detection performance of serum free light chain (sFLC) in two platforms and evaluate the comparability of serum free light chain results in patients with multiple myeloma (MM).Methods:To evaluate the detection performance (repeatability, accuracy, linear range, reference range, interfering substances, etc.) of sFLC kit based on polyclonal antibodies. Spearman correlation analysis and Bland-Altman were used to analyze 214 sFLC results obtained on two detection platforms at the same time to evaluate the correlation between the results of the two methods and analyze the causes of methodological bias. 119 cases with aMM and 23 cases of disease control group (AL, WM, POEMS syndrome, MGUS, diffuse large B-cell lymphoma) initially diagnosed in the hematology department of Zhongshan Hospital of Fudan University from March 2020 to March 2021 were all included. A retrospective analysis was conducted to calculate the area under the curve of receiver operating characteristic (AUC-ROC) and obtain the optimal sensitivity and specificity cut-off points for the diagnosis of MM patients on monoclonal antibody platform.Results:Repeatability, accuracy, linear range, reference interval and anti-interfering capacity of the detection platform based on polyclonal antibodies were verified to meet clinical needs. The overall consistency of FLC/κ, FLC/λ and κ/λ ratios in two methods was 89.3%, 84.1% and 77.1% respectively; but the correlation results were highly heterogeneous. The correlation coefficient of FLC/κ R 2 was 0.922( P<0.001), while the correlation coefficients R 2of the FLC/λ and κ/λ ratios were only 0.349 and 0.441( P<0.001). After segment analysis, it was found that the correlation of FLC/λ was improved within the linear range and R 2 could rise to 0.78( P<0.001). Compared with monoclonal antibody platform, the vast majority points of FLC/κ fell within the 95% limit by Bland Altman analysis. While the results of FLC/λ on polyclonal antibody platform showed significant positive bias. The AUC of MM diagnosis on monoclonal antibody platform was 0.751 ( P=0.001), and the optimal cutoff value was 24.67. Conclusion:The overall consistency between the two platforms was good, but there were significant differences between the results, so they were not comparable and could not be interchanged. For monitoring the prognosis of patients with multiple myeloma, the same platform should be selected for testing.

The global COVID-19 coronavirus pandemic has infected over 109 million people, leading to over 2 million deaths up to date and still lacking of effective drugs for patient treatment. Here, we screened about 1.8 million small molecules against the main protease (M^pro) and papain like protease (PL^pro), two major proteases in severe acute respiratory syndrome-coronavirus 2 genome, and identified 1851M^pro inhibitors and 205 PL^pro inhibitors with low nmol/l activity of the best hits. Among these inhibitors, eight small molecules showed dual inhibition effects on both M^pro and PL^pro, exhibiting potential as better candidates for COVID-19 treatment. The best inhibitors of each protease were tested in antiviral assay, with over 40% of M^pro inhibitors and over 20% of PL^pro inhibitors showing high potency in viral inhibition with low cytotoxicity. The X-ray crystal structure of SARS-CoV-2 M^pro in complex with its potent inhibitor 4a was determined at 1.8 Å resolution. Together with docking assays, our results provide a comprehensive resource for future research on anti-SARS-CoV-2 drug development.
Humans ; Antiviral Agents/chemistry* ; COVID-19 ; COVID-19 Drug Treatment ; High-Throughput Screening Assays ; Molecular Docking Simulation ; Protease Inhibitors/chemistry* ; SARS-CoV-2/enzymology* ; Viral Nonstructural Proteins

Objective: To review the results of inter-laboratory comparisons in Shanghai glycohemoglobin harmonization program from 2010 to 2018, and to analyze the evolution of quality levels of HbA_1c determination, so as to provide the reference for improving the HbA_1c determination quality in China. Methods: Retrospective analysis. The comparison data of Shanghai Glycohemoglobin Harmonization Program from 2010 to 2018 was collected. And the change trend was analyzed about hospital and determination method distribution. The judgment criteria, quarterly and annual pass rate, bias and coefficient of variation of the results of the inter-laboratory comparison were analyzed retrospectively, and the results were compared with the results of External Quality Assessment Programme carried out by the National Center for Clinical Laboratories, Shanghai Center for Clinical Laboratories and College of American Pathologists (CAP). The data in the first quarter of 2019 was collected and the imprecision, bias and sigma were calculated, which were drew in the evaluation model of sigma combined with biomedical variation parameters. Results: The number of participating laboratories increased from 9 in Shanghai to 192 in the whole country, with an average annual growth rate of 76.6%. The quarterly comparison criteria improved from ±8% to ±6% and the passing rate of participating laboratories increased from 39.1% to nearly 90%. The maximum CV of each instrument among laboratories decreased from 14.3% to 4.8%. In the first quarter of 2019, nearly 60% of the laboratories met 6σcriteria and more than 95% of the laboratories met the "standard criteria" in the model of biological variation parameters. Conclusion Shanghai Glycohemoglobin harmonization program has improved the harmonization of HbA_1c test results among the participating laboratories.

Objective:To establish autoverification rules for coagulation tests in multicenter cooperative units, in order to reduce workload for manual review of suspected results and shorten turnaround time (TAT) of test reports, while ensure the accuracy of results.Methods:A total of 14 394 blood samples were collected from fourteen hospitals during December 2019 to March 2020. These samples included: Rules Establishment Group 11 230 cases, including 1 182 cases for Delta check rules; Rules Validation Group 3 164 cases, including 487cases for Delta check; Clinical Application Trial Group 77 269 cases. Samples were analyzed for coagulation tests using Sysmex CS series automatic coagulation analyzers, and the clinical information, instrument parameters, test results, clinical diagnosis, medication history of anticoagulant and other relative results such as HCT, TG, TBIL, DBIL were summarized; on the basis of historical data, the 2.5 and 97.5 percentile of all data arranged from low to high were initially accumulated; on the basis of clinical suggestions, critical values and specific drug use as well as relative guidelines, autoverification rules and limits were established.The rules were then input into middleware, in which Stage I/Stage II validation was done. Positive coincidence, negative coincidence, false negative, false positive, autoverification pass rate, passing accuracy (coincidence of autoverification and manual verification) were calculated. Autoverification rules underwent trial application in coagulation results reports.Results:(1) The autoverification algorisms involve 33 rules regarding PT/INR, APTT, FBG, D-dimer, FDP,Delta check, reaction curve and sample abnormalities; (2)Autoverification Establishment Group showed autoverification pass rate was 68.42% (7 684/11 230), the false negative rate was 0%(0/11230), coincidence of autoverification and manual verification was 98.51%(11 063/11 230), in which positive coincidence and negative coincidence were respectively 30.09% (3 379/11 230) and 68.42%(7 684/11 230); Autoverification Validation Group showed autoverification pass rate was 60.37%(1 910/3 164), the false negative rate was 0%(0/11 230), coincidence of autoverification and manual verification was 97.79%(3 094/3 164), in which positive coincidence and negative coincidence were respectively 37.42%(1 184/3 164) and 60.37%(1 910/3 164); (3) Trialed implementation of these autoverification rules on 77 269 coagulation samples showed that the average TAT shortened by 8.5 min-83.1 min.Conclusions:This study established 33 autoverification rules in coagulation tests. Validation showedthese rules could ensure test quality while shortening TAT and lighten manual workload.

Objective To study the clinical value and the safety of endoscopic retrograde cholangio-pancreatography(ERCP)for chronic pancreatitis in the pediatric population.Methods Clinical data,endo-scopic reports,and radiography of chronic pancreatic patients aged from 2 to 17 years old who underwent ERCP between Jan.1,2008 and Apr.30,2014 at Endoscopy Center of Ruijin Hospital were reviewed.These patients were divided into 3 groups,patients aged from 2 to 6 years old as children group(n =27),patients aged from 7 to 12 years old as juvenile group(n =35)and patients aged from 13 to 17 as adolescent group(n =25). Their clinical data and complications were analyzed by groups.Results A total of 170 ERCP procedures were performed in 87 patients,with the success rate of 99.4%(169 /170).All patients were diagnosed as having chronic pancreatitis by ERCP.Success rates of children group,juvenile group and adolescent group were 97.6%(40 /41),100.0%(72 /72)and 100.0%(57 /57)respectively.There was no statistically significance among the three groups(P =0.207).No serious complications like bleeding or perforation occurred.No proce-dure-related mortality occurred in this study either.The rate of post-ERCP adverse events in children group, juvenile group and adolescent group were 41.5%(17 /41),25.0%(18 /72)and 24.6%(14/57),respective-ly.Children group had more adverse events than the juvenile group(P =0.039)and adolescent group(P =0.045).Conclusion Pediatric ERCP is highly effective in the pediatric population with chronic pancreatitis. The incidence of post ERCP adverse events is higher among the youngest children.

Objective To compare the clinical choosing principles of sedated colonoscopy with con-ventional colonoscopy. Methods Outpatients who were willing to accept colonoscopy with or without seda-tion were prospectively recruited,which were assigned to sedated colonoscopy group(n=362)and conven-tional colonoscopy group(n=323). All patients and endoscopists were asked to answer a self-administered questionnaire. The colonoscopy completion,operation time,procedure-related discomfort,and questionnaire results of the two groups were compared and statistically analyzed. Results The completion rate was 98. 9%in the sedated colonoscopy group(358/362)and 89. 8% in the conventional colonoscopy group(290/323) ( P=0. 337 ). The operation time of sedated and conventional group were( 5. 60 ± 3. 25 ) minutes and (7. 71 ± 5. 70)minutes respectively(P<0. 001). And the average cost was CNY 886. 54 per patient in se-dated group and CNY 386. 00 per patient in the conventional group. Patient satisfaction score of conventional group and sedated group were 4(3-4)and 3(2-3)points(P<0. 001),while endoscopist satisfaction score was 4(3-4)and 4(4-4)(P<0. 001). A total of 354 patients(97. 79%)in the sedated group and 225 pa-tients(69. 66%)in the conventional group showed willingness to repeat the identical colonoscopy( P <0. 001). Patients who were male(P=0. 035),having no past abdominal operations(P<0. 001),or no ab-dominal pain during colonoscopy( P =0. 015 )in the conventional group preferred to repeat conventional colonoscopy. Conclusion Although the examination time of conventional colonoscopy is longer than sedated colonoscopy,it could reduce anesthesia risk and the cost. Conventional colonoscopy remains an irreplaceable examination of colorectal diseases in developing countries. Physicians should not only focus on patients'com-fort during endoscopy,but also help patients make a decision based on their actual situation and endoscopic indications to make the best of medical resources.

Objective To investigate the clinical value of quality intima-media thicknes (QIMT),quality arterial stiffness(QAS) and XStrain in assessing the ventricular-arterial coupling (VAC) in patients with uremia.Methods Sixty-five patients with uremia and 30 normal subjects were enrolled in this study as the uremic group and control group respectively.Ultrasound examination for the cardiac and carotid artery was performed and some parameters were obtained,such as left ventricular ejection fraction(LVEF),E/e,Tei index,stiffness (β),compliance coefficinet (CC),pulse wave velocity (PWV),intima-media thicknes (IMT),strain,and so on.The sum of left ventricular systolic radial strain and carotid arterial diastolic radial strain was used as a new parameter (VACs) for assessing VAC.The correlation between VACs and VACv,a traditional method to evaluate VAC,was analyzed.The parameters obtained by ultrasonic techniques were compared between two groups.The intra-and inter-observer reliability of XStrain was assessed by intraclass correlation coefficient.The multiple linear regression and ROC curve were used to analyze the independent factor of cardiovascular dysfunction.Results ①The pulse pressure,E/e,Tei index,as well as β,PWV and IMT were larger in uremic group than control group significantly (P ＜0.05).②The function of VAC was decreased in uremic group,and the VACv and VACs were lower than control group significantly (P ＜0.05).③There was strong positive relation between VACs and VACv (r =0.908,P =0.000).The reproducibility of XStrain technique was well.④VACs,β,VACv and PWV could be considered as independent factor of cardiovascular dysfunction and performance of VACs was the largest (standardized coefficient was-0.582).A cutoff value of VACs for assessing cardiovascular dysfunction was less than 23.60,which had both higher sensitivity and specificity (96.4％,81.1％).Conclusions The QIMT,QAS and XStrain can be used to assess cardiovascular structure,function and VAC accurately and globally,which can be consider as an effective method for detecting cardiovascular complication and improving prognosis of uremic patients.

Objective To evaluate the value of urinary liver-type fatty acid binding protein (L-FABP)as a biomarker in prediction of renal function progression in patients with chronic glomerulonephritis (CGN). Methods A total of 123 patients with newly diagnosed CGN by renal biopsy in Shanghai Renji Hospital between 2004 January and 2005 December were enrolled in the study,Twenty-eight healthy subjects were used as control group.Urine samples were collected before biopsy and treatment,and urinary L-FABP was measured by ELISA.The patients with follow-up every three months for 5 years were divided into progressive group and nonprogressive group.The progression of kidney function impairment was defined as a reduction of GFR ≥ 5 ml·min-1·(1.73 m2)-1·year-1 during follow-up.The risk factors of progressive renal function were evaluated and the Spearman correlation analysis was performed to find out the prognostic indicator of renal function deterioration. Results Urinary L-FABP level of CGN patients was significantly higher than that of healthy control group (P＜0.01).Urinary L-FABP in CGN patients was negatively correlated with eGFR (r=-0.565,P＜0.01) and positively correhted with proteinuria (r=0.501,P＜0.01) and Scr (r=0.601,P＜0.01).Kaplan-Meier analysis showed that urinary L-FABP excretion＞76.58 μg/g·cr predicted progression of renal function.The AUC of urinary L-FABP for prognosis of CGN progression was 0.95,with 87.5％ of sensitivity and 90.5％of specificity at the cutoff value of 119.8 μg/g·cr,which revealed its great value of predicting the prognosis of CGN patients. Conclusion Urinary L-FABP can be a novel biomarker of evaluation for renal injury and early progressive renal function deterioration in patients with CGN.

The graft-versus-tumor (GVT) effect of T cells induced by tumor antigen-pulsed CD8α(+) dendritic cells (DCs) in vitro was investigated in this study. Immature CD8α(+) DCs were prepared from C57BL/6 (H-2(b)) bone marrow cells by using a cytokine cocktail. On the 3rd day of culture, CD8α(+) DCs were pulsed by allogeneic (Balb/c, H-2(d)) EL9611 leukemia antigen, or RM-1 syngeneic prostate cancer antigen, with the concentration series of 0, 2.5, 5.0, 10.0, 20.0 μg/mL, respectively, then antigen-loaded immature CD8α(+) DCs were co-cultured with syngeneic T cells according to the DC/T ratio of 1:1, 2:1 and 4:1. T cell proliferation was measured by MTT assay. Cytokines including interferon gamma (IFN-γ) and interleukin-10 (IL-10) in CD8α(+) DCs and T co-culture supernatant were detected by using ELISA. Cytotoxic effect of antigen-specific T cells was tested by LDH release assay. Conventional mature DCs (mDCs) induced from C57BL/6 (H-2(b)) bone marrow cells by using granulocyte-macrophage colony stimulating factor (GM-CSF) and interleukin-4 (IL-4) served as a control. The results showed that the proliferative activity of T cells stimulated by CD8α(+) DCs loaded with allogeneic or syngeneic tumor antigen was augmented with the CD8α(+) DC/T ratio increased (P<0.05). When antigen concentration ≤ 5 μg/mL and CD8α(+) DC/T ratio ≤ 2:1, the ability of CD8α(+) DCs to stimulate T cell proliferation was higher than mDC control in allogeneic tumor antigen-pulsed groups (P<0.05), but not in syngeneic tumor antigen-pulsed groups (P>0.05). The level of IFN-γ and IL-10 in CD8α(+) DCs and T cell co-culture supernatant were increased in both allogeneic and syngeneic antigen-pulsed groups (P<0.05), and the cytokine level was higher in allogeneic antigen-pulsed groups than in syngeneic antigen groups when the CD8α(+) DC/T was 1:1 or 2:1 (P<0.05). There existed a negative correlation between the level of IL-10 and T cell proliferation. T cell cytotoxicity assay showed that when CD8α(+) DCs were pulsed with allogeneic tumor antigen, the maximal T cell killing efficiency could reach (100±7.7)%, whereas syngeneic tumor antigen-pulsed group had only (65.0±3.4)%. It was concluded that syngeneic and allogeneic tumor antigen-pulsed immature CD8α(+) DCs could stimulate T cells to exert the GVT effect in vitro, and the GVT effect was more obvious with allogeneic tumor antigen than with syngeneic tumor antigen. The optimal condition was low allogeneic tumor antigen pulsation (≤ 5 μg/mL) and low CD8α(+) DC/T ratio (1:1 and 2:1).

The graft-versus-tumor (GVT) effect of T cells induced by tumor antigen-pulsed CD8α+dendritic cells (DCs) in vitro was investigated in this study.Immature CD8α+ DCs were prepared from C57BL/6 (H-2b) bone marrow cells by using a cytokine cocktail.On the 3rd day of culture,CD8α- DCs were pulsed by allogeneic (Balb/c,H-2d) EL9611 leukemia antigen,or RM-1 syngeneic prostate cancer antigen,with the concentration series of 0,2.5,5.0,10.0,20.0 μg/mL,respectively,then antigen-loaded immature CD8α+ DCs were co-cultured with syngeneic T cells according to the DC/T ratio of 1∶1,2∶1and 4∶1.T cell proliferation was measured by MTT assay.Cytokines including interferon gamma (IFN-γ)and interleukin-10 (IL-10) in CD8α+ DCs and T co-culture supernatant were detected by using ELISA.Cytotoxic effect of antigen-specific T cells was tested by LDH release assay.Conventional mature DCs　(mDCs) induced from C57BL/6 (H-2b) bone marrow cells by using granulocyte-macrophage colony　stimulating factor (GM-CSF) and interleukin-4 (IL-4) served as a control.The results showed that the　proliferative activity of T cells stimulated by CD8α+ DCs loaded with allogeneic or syngeneic tumor antigen was augmented with the CD8α+ DC/T ratio increased (P＜0.05).When antigen concentration ≤ 5μg/mL and CD8α+ DC/T ratio ≤ 2∶1,the ability of CD8α+ DCs to stimulate T cell proliferation was　higher than mDC control in allogeneic tumor antigen-pulsed groups (P＜0.05),but not in syngeneic tumor antigen-pulsed groups (P＞0.05).The level of IFN-γ and IL-10 in CD8α+DCs and T cell co-culture　supernatant were increased in both allogeneic and syngeneic antigen-pulsed groups (P＜0.05),and the　cytokine level was higher in allogeneic antigen-pulsed groups than in syngeneic antigen groups when　the CD8α+DC/T was 1∶1 or 2∶1 (P＜0.05).There existed a negative correlation between the level of　IL-10 and T cell proliferation.T cell cytotoxicity assay showed that when CD8α+ DCs were pulsed with　allogeneic tumor antigen,the maximal T cell killing efficiency could reach (100±7.7)％,whereas syngeneic tumor antigen-pulsed group had only (65.0±3.4)％.It was concluded that syngeneic and allogeneic tumor antigen-pulsed immature CD8α+ DCs could stimulate T cells to exert the GVT effect in vitro,and the GVT effect was more obvious with allogeneic tumor antigen than with syngeneic tumor antigen.The optimal condition was low allogeneic tumor antigen pulsation (≤ 5 μg/mL) and low CD8α+ DC/T　ratio (1∶1 and 2∶1).