Objective:To investigate the optimal delay time in the quantitative assessment of myocardial fibrosis based on dual-energy CT extracellular volume fraction (DECT-ECV), using MRI as a reference.Methods:Thirty patients with confirmed or suspected of cardiomyopathy were prospectively enrolled in this study. All the patients underwent both cardiac DECT and MRI examination within one week. According to the imaging features of late gadolinium enhancement (LGE) on MRI, myocardial segments were classified into 3 types: ischemic LGE segments, non-ischemic LGE segments and negative LGE segments. According to the DECT delay time, the whole and segmental myocardium were divided into 3 groups: delay of 3 min (Group A), delay of 5 min (Group B) and delay of 7 min (Group C). Correlation and agreement between CT-ECV and MRI-ECV were performed on a basis of overall myocardium and segmental myocardium. Pearson or Spearman test was used for correlation analysis and Bland-Altman test was used for consistency analysis.Results:Thirty patients with 480 segments were finally included in our study. In the analysis based on overall myocardium, MRI-ECV was 33.12%±4.29%, and CT-ECV were 35.81%±4.48%, 36.02%±4.56%, and 36.58%±4.69% in Group A, B, and C, respectively. The agreement between DECT-ECV and MRI-ECV results was good, with the correlation coefficients of 0.878 (group A), 0.955 (Group B) and 0.947 (Group C) (all P<0.001). In the analysis based on segmental myocardium, as for the ischemic LGE myocardial segments, MRI-ECV was 34.60%(31.70%,39.40%), and CT-ECV were 37.50 (34.20, 41.90), 38.20%(36.20%, 40.60%)and 39.40%(35.50%,42.40%)in Group A, B, and C, respectively. The agreement between DECT-ECV and MRI-ECV results was good, with the correlation coefficients of 0.559, 0.695 and 0.682 (all P<0.001) for groups A, B and C, and as for non-ischemic LGE myocardial segments, MRI-ECV was 35.10% (32.68%, 38.70%), and CT-ECV were 38.15% (35.13%, 41.75%), 39.25% (35.78%, 42.20%) and 39.60% (35.88%,42.90%) in Group A, B, and C. The correlation coefficients of CMR-ECV and DECT-ECV of groups A, B and C were 0.531, 0.772 and 0.744 (all P<0.001), showing good agreement; as for negative LGE myocardial segments, MRI-ECV and CT-ECV of Group A, Group B, Group C were 28.50%(27.00%, 30.10%), 31.10%(28.70%, 34.60%), 31.30%(28.40%, 33.80%), 31.30%(29.20%, 34.80%). The correlation coefficients between MRI-ECV and DECT-ECV of group A, B and C were 0.273, 0.508 and 0.425 (all P<0.001), which also showed good agreement. Conclusions:DECT-ECV can be used for quantitative evaluation of myocardial histological features. DECT-ECV with a 5 min and 7 min delay shows good correlation and agreement with MRI-ECV. In order to make this technology more well-known and improve its application capability, our recommendation for clinical practice is a 5 min delay after contrast administration in clinical practice.

ObjectiveTo study the traditional Chinese medicine （TCM） syndromes of children with alopecia areata， and provide evidence for TCM differentiation and treatment in clinic. MethodsA retrospective analysis was conducted on the clinical data of 800 children with alopecia areata admitted to the Hair Medicine Center of the China-Japan Friendship Hospital from January 1， 2012 to December 31， 2021. The clinical data of the children were collected using a four-examination information questionnaire， including clinical characteristics （age of consultation， age of onset， course of disease， family history， severity grading）， alopecia areata-related factors （triggers）， and four-examination information （including sleep， diet， emotions， bladder and bowel function， etc.）. Descriptive frequency analyses， rank sum tests， factor analyses and cluster analyses were performed， and the distribution of the major TCM syndromes was summarised with the clinical data. ResultsThere were 800 children with alopecia areata， including 449 males and 351 females； 8 cases （1.00%） were in infancy， 36 cases （4.50%） were in early childhood， 180 cases （22.50%） were in preschool， 380 cases （47.50%） were in school age， and 196 cases （24.50%） were in puberty at the time of consultation； the average age of consultation was 8.31±3.86 years， the average age of onset of disease was 5.40±3.82 years， and the average duration of disease was 2.94±2.77 years； 527 children （65.87%） with severe alopecia areata； 85 children （13.56%） had a family history of alopecia areata； 772 children （96.50%） had unknown triggers for their first alopecia areata， and 28 children （3.50%） reported the presence of obvious triggers， including fright （9 cases）， high fever （5 cases）， allergic reactions （4 cases）， micronutrient （zinc， iron， etc.） deficiencies （4 cases）， inappropriate diet （2 cases）， environmental factors （1 case， new house renovation）， atopic dermatitis （1 case）， atopic asthma （1 case）， and pneumonia （1 case）. A total of 40 four-examination information items were collected， among which the frequency of kicking quilts was the highest with 380 cases （47.50%）， followed by picky eating （369 cases， 46.13%）， sleeplessness （334 cases， 41.75%）， irritability （334 cases， 41.75%）， partiality towards certain foods （306 cases， 38.25%）， impulsiveness （297 cases， 37.13%）， dry stools （233 cases， 29.13%）， yellow urine （215 cases， 26.88%）， nail biting （213 cases， 26.63%）， bad breath （211 cases， 26.38%）. According to factor analysis and cluster analysis， five types of TCM syndromes were obtained， in order as qi and blood deficiency syndrome （110 cases， 13.75%）， spleen deficiency syndrome （114 cases， 14.25%）， kidney essence deficiency syndrome （140 cases， 17.50%）， dietary stagnation syndrome （150 cases， 18.75%）， and liver depression and spleen deficiency syndrome （286 cases， 35.75%）. Patients in each age group and SALT grading are mainly liver depression and spleen deficiency syndrome. ConclusionThe TCM symptoms of children with alopecia areata are mainly based on qi and blood deficiency syndrome， spleen deficiency syndrome， kidney essence deficiency syndrome， dietary stagnation syndrome， and liver depression and spleen deficiency syndrome， of which liver depression and spleen deficiency syndrome is the most common type at different ages and stages of the disease.

Objective To generate and identify the Itgbl1(integrin beta-like)promoter-driven Cre knock-in mouse line.Methods Itgbll-Cre knock-in mice were generated using clustered regularly interspaced short palindromic repeats(CRISPR)/CRISPR-associated protein 9(Cas9)gene editing.The Itgbl1-Cre mice were crossed with the Cre reporter ROSALSL-tdTomato)mice to detect the expression profile of Cre activity.The tdTomato expression pattern across tissues and cell-specific markers were used to identify the cell types of Itgbl1-expressing cells and their progeny.Results and Conclusion tdTomato was specifically expressed in mucous acinar cells of the sublingual gland,pancreatic islet cells,and gastric endocrine cells.In addition,tdTomato expression was also found in some of the neurons of the retina and brain,as well as in a few cells in the serosal layer of the intestine,articular cartilage,periosteum,and bone marrow.The first Itgbl1-Cre recombinase transgenic mouse line was established,which can specifically label the mucous acinar cells of the sublingual gland.

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 investigate the clinical efficacy of elderly patients with diffuse large B-cell lymphoma (DLBCL) and the influencing factors of prognosis.Methods:The clinical data of 76 elderly (≥60 years old) patients with DLBCL admitted to Huadong Hospital Affiliated to Fudan University between January 2015 and December 2019 were retrospectively analyzed. The R-CHOP regimen was the preferred treatment for 54 patients, while the remaining patients received R-miniCHOP, CHOP or other regimens or supportive treatments due to age, physical condition, economic factors, etc., which were not included in the efficacy analysis. Kaplan-Meier method was used to analyze the survival status of patients. Multivariate Cox proportional risk model was used to analyze the prognostic factors.Results:Among the 54 patients who preferred R-CHOP regimen for treatment, 26 cases (48.1%) achieved complete remission and 14 cases (25.9%) achieved partial remission, and the total effective rate was 74.1% (40/54); Among them, the total effective rate of 37 cases aged 60-69 years was 70.3% (26/37), and the total effective rate of 17 cases aged 70-79 years was 82.4% (14/17); there was no statistically significant difference in the total effective rate between the two groups ( χ2 = 3.01, P = 0.390). All 76 patients were followed up for 1-60 months. As of the last follow-up, 49 patients (64.5%) died, with the median overall survival (OS) time of 16 months and 5-year OS rate of 35.5%. Kaplan-Meier method showed that age ≥ 70 years old at initial diagnosis, Eastern Cooperative Oncology Group (ECOG) score ≥ 2 points, presence of B symptoms, international prognosis index (IPI) score >3 points, elevated lactate dehydrogenase, immunohistochemistry positive for bcl-2, and non-germinal center type were associated with poor OS (all P < 0.05). Multivariate Cox analysis showed that age ≥ 70 years old at initial diagnosis, presence of B symptoms, positive expression of bcl-2, non-germinal center type were independent risk factors for OS (all P < 0.05). Conclusions:Elderly DLBCL patients have poor survival. Old age at initial diagnosis, B symptoms, bcl-2 positive, and non-germinal center type are independent risk factors of prognosis.

Objective:To analyze the differences in neuronal activation during fear memory extinction in various sub-regions of the hippocampus in post-traumatic stress disorder(PTSD)mice.Methods:Two immediate early gene-pro-tein labeling strategies were employed to label neurons associated with fear extinction in PTSD mice.In the first group,Arc protein in hippocampal neurons was labeled and observed through immunofluorescence staining in wild-type mice.In the second group,Fos-CreERT2;Ai9 transgenic mice were injected with tamoxifen 23 hours prior to inducing fear memory extinction,and the relevant neurons were labeled with fluorescent proteins for observation.The number of labeled hippocampal neurons and the dendritic branch structure were analyzed to compare the activation levels of hipp-ocampal neurons and the plasticity of neuronal dendrites.Results:The two groups of Arc and Fos positive neurons were mainly distributed in the dorsal hippocampus,in which Arc protein chromogenic was enriched in CA3 and DG subre-gions,while CA1 and CA2 subregions were scattered,while Fos-positive neurons were enriched in the DG subregion of hippocampus and scattered in CA1,CA2 and CA3 subregions.Compared to the control group,there was no significant difference in the number of neurons expressing Arc protein in each subregion of the hippocampus in the PTSD group.The number of Fos-positive neurons in CA1,CA3 and DG subregions in the hippocampus of the PTSD group was signifi-cantly increased(P＜0.01).The dendritic branches of neurons in the hippocampal region were observed and analyzed in Fos-CreERT2;Ai9 mice from both groups,but no significant changes were found.Conclusion:Abnormal activation of neurons occurs in different subregions of the hippocampus during fear extinction in PTSD mice,although there are no significant plasticity changes in the dendritic branches of the activated neurons.

Somatostatin (SST) is an inhibitory polypeptide hormone that plays an important role in a variety of biological processes. Somatostatin receptor 2 (SSTR2) is the most widely expressed somatostatin receptor. However, the specific cell types expressing Sstr2 in the tissues have not been investigated. In this study, we detected the expression pattern of SSTR2 protein in mouse at different development stages, including the embryonic 15.5 days and the postnatal 1, 7, 15 days as well as 3 and 6 months, by multicolour immunofluorescence analyses. We found that Sstr2 was expressed in some specific cells types of several tissues, including the neuronal cells and astrocytes in the brain, the mesenchymal cells, the hematopoietic cells, the early hematopoietic stem cells, and the B cells in the bone marrow, the macrophages, the type Ⅱ alveolar epithelial cells, and the airway ciliated cells in the lung, the epithelial cells and the neuronal cells in the intestine, the hair follicle cells, the gastric epithelial cells, the hematopoietic stem cells and the nerve fibre in the spleen, and the tubular epithelial cells in the kidney. This study identified the specific cell types expressing Sstr2 in mouse at different developmental stages, providing new insights into the physiological function of SST and SSTR2 in several cell types.
Mice ; Animals ; Receptors, Somatostatin/metabolism* ; Hematopoietic Stem Cells/metabolism* ; Epithelial Cells

Objective To investigate the association between baseline IgM level and treatment response to ursodeoxycholic acid (UDCA) in patients with primary biliary cholangitis (PBC). Methods A retrospective analysis was performed for the clinical data of 637 PBC patients who were diagnosed and treated with UDCA for the first time in The Fifth Medical Center of Chinese PLA General Hospital from January 2010 to January 2020. The PBC patients were divided into UDCA complete response group with 436 patients and UDCA poor response group with 201 patients, and baseline clinical data were compared between the two groups. According to the optimal cut-off value of IgM determined by the area under the ROC curve (AUC) of baseline indices in predicting the risk of poor treatment response, the patients were divided into IgM ≥1.5×ULN group and IgM < 1.5×ULN group, and baseline parameters, treatment response, and prognostic model score were compared between groups. The t -test was used for comparison of normally distributed continuous data between two groups, and the Mann-Whitney U test was used for comparison of non-normally distributed continuous data between two groups; the chi-square test was used for comparison of categorical data between two groups. The Cochran-Mantel-Haenszel test was used for subgroup analysis, and forest plots were plotted for related risk values. Results Compared with the UDCA complete response group, the UDCA poor response group had significantly higher proportion of patients with liver cirrhosis, levels of total bilirubin, aspartate aminotransferase (AST), alkaline phosphatase (ALP), total bile acid, total cholesterol (TC), IgA, and IgM, and positive rate of anti-Gp210 antibody at baseline ( χ 2 =4.596, Z =-9.932, -8.931, -8.361, -7.836, -4.694, -3.242, and -2.115, χ 2 =15.931, all P < 0.05). The UDCA poor response group had significantly higher Mayo Risk Score, Globe score, and UK-PBC risk score than the UDCA complete response group ( t =4.092, Z =-10.910 and -11.646, all P < 0.001). Compared with the normal IgM group, the elevated IgM group had significantly higher levels of AST, ALP, TC, IgA, and IgG and a significantly higher positive rate of anti-Gp210 antibody ( Z =-3.774, -5.063, -4.344, -2.051, and -6.144, χ 2 =25.180, all P < 0.05). IgM had an AUC of 0.552 in predicting poor treatment response. Compared with the IgM < 1.5×ULN group, the IgM ≥1.5×ULN group had significantly higher levels of AST, ALP, TC, and IgG, a significantly higher positive rate of anti-Gp210 antibody, and a significantly higher poor UDCA response rate ( Z =-4.193, -5.044, -3.250, and -5.465, χ 2 =25.204 and 8.948, all P < 0.05). IgM ≥1.5×ULN had an odds ratio of 1.416 (95% confidence interval [ CI ]: 1.129-1.776, P =0.003) in predicting poor response. The subgroup analysis showed that for patients without liver cirrhosis, IgM ≥1.5×ULN had an odds ratio of 1.821 (95% CI : 1.224-2.711, P =0.003) in predicting poor response. Conclusion Baseline IgM level has an important value in predicting UDCA response. IgM level should be closely monitored during treatment in PBC patients with a high baseline IgM level, and second-line drugs should be given in time if the abnormality persists.

Onset with fever and back pain, an 81-year-old man had sudden oliguria and progressively elevated serum creatine from normal range to 660 μmol/L within 1 week after receiving contrast agents, various antibiotics, and several nonsteroidal anti-inflammation drugs. Urine output recovered after supportive treatment. However, his serum creatinine level rose again soon after a temporary decline accompanied by gross hematuria with almost normal morphology, nephrotic proteinuria, and hypoalbuminemia. Renal biopsy revealed necrotizing glomerulonephritis. Methylprednisolone was intravenously administrated 500 mg per day for 3 days, followed by oral glucocorticoids and cyclophosphamide. Gradually the patient′s serum creatinine descended to 144 μmol/L.

Objective:To systematically evaluate the efficacy and safety of demethylation drugs intreatment of myelodysplastic syndrome (MDS), and to provide reliable basis and guidance for clinical application of these drugs.Methods:The randomized controlled trials (RCT) of demethylation drugs for treatment of MDS published in the PubMed, Web of Science, Embase, Cochrane Library, Chinese Journal Full-text Database (CNKI), Chinese Biomedical Literature Database (CBM) and VIP database from January 2000 to December 2019 were searched by computer. RevMan 5.3 software was used for Meta-analysis of the efficacy and safety.Results:Seven RCT studies involving 1 172 patients were obtained. Meta-analysis showed that the complete remission rate ( OR = 6.26, 95% CI 1.74-22.49, P = 0.005), partial remission rate ( OR = 4.65, 95% CI 1.51-14.29, P = 0.007), overall response rate ( OR = 14.14, 95% CI 7.27-27.51, P < 0.01), hematology improves ( OR = 3.47, 95% CI 1.44-8.32, P = 0.005) in the demethylation drug treatment group were better than those in the best supportive treatment group. Meanwhile, the overall survival of patients in azacytidine group was improved ( HR = 0.62, 95% CI 0.50-0.77, P < 0.01). In terms of adverse reactions, demethylation drugs increased the incidence of neutropenic fever ( OR = 4.19, 95% CI 2.26-7.76, P < 0.01). However, there was no significant difference in the incidence of neutropenia, thrombocytopenia, anemia, infection, nausea, liver damage and fatigue between the two groups (all P > 0.05). Conclusions:The effect of demethylation drugs in treatment of MDS is obvious, the remission rate can be improved, and azacytidine can prolong the patients' survival, but demethylation drugs increase the incidence of neutropenia fever. In the clinical application of demethylation drugs, it is necessary to further carefully evaluate their clinical safety.