Objective To predict the core targets and action pathways of Hedysari Radix based on UPLC-MS/MS and network pharmacology methods,and to verify the results of network pharmacology by molecular docking and molecular dynamics techniques.This article aims to investigate immune regulation mechanism of effective components absorbed into blood from Hedysari Radix.Methods Qualitative quantification of effective components absorbed into blood from Hedysari Radix were operated by using UPLC-MS/MS technique.The corresponding targets of effective components absorbed into blood from Hedysari Radix were screened by TCMSP and HERB databases.Targets of immune-related disease were obtained through DisGeNET,OMIM,TTD,and MalaCards databases.The network of"components absorbed into blood from Hedysari Radix-immune-related diseases"was then constructed.GO and KEGG enrichment analysis and mapped the PPI network were performed.Molecular docking and molecular dynamics techniques were applied for validation.Results A total of 8 prototype components absorbed into blood,synergistically acting on 101 targets,were identified by UPLC-MS/MS.They mediated 538 biological processes including immune response,positive regulation of gene expression,receptor binding,and cytokine activity.Meanuhile,116 signaling pathways,such as HIF-1,Toll-like receptor,JAK-STAT,T cell receptor,PI3K-Akt,and FoxO etc.were involved.The core targets were MAPK14,PTGS2,MMP9,PPARG,CCND1,etc..The results of molecular docking showed that formononetin and calycosin had strong docking binding activity with MAPK14.And molecular dynamics simulations further demonstrated that the binding between MAPK14 and formononetin or calycosin had good structural stability and binding affinity.Conclusion The results of serum pharmacochemistry,network pharmacology and molecular dynamics were verified to reveal the material basis and mechanism of Hedysari Radix in regulating immunity.The aim of this study is to provide scientific basis for its immunomodulatory mechanism.

BACKGROUND: Previous research demonstrated that a homozygous mutation of g.136372044G>A (S12N) in caspase recruitment domain family member 9 ( CARD9 ) is critical for producing Aspergillus fumigatus -induced ( Af -induced) T helper 2 (T H 2)-mediated responses in allergic bronchopulmonary aspergillosis (ABPA). However, it remains unclear whether the CARD9S12N mutation, especially the heterozygous occurrence, predisposes the host to ABPA. METHODS: A total of 61 ABPA patients and 264 controls (including 156 healthy controls and 108 asthma patients) were recruited for sequencing the CARD9 locus to clarify whether patients with this heterozygous single-nucleotide polymorphisms are predisposed to the development of ABPA. A series of in vivo and in vitro experiments, such as quantitative real-time polymerase chain reaction, flow cytometry, and RNA isolation and quantification, were used to illuminate the involved mechanism of the disease. RESULTS: The presence of the p.S12N mutation was associated with a significant risk of ABPA in ABPA patients when compared with healthy controls and asthma patients, regardless of Aspergillus sensitivity. Relative to healthy controls without relevant allergies, the mutation of p.S12N was associated with a significant risk of ABPA (OR: 2.69 and 4.17 for GA and AA genotypes, P = 0.003 and 0.029, respectively). Compared with patients with asthma, ABPA patients had a significantly higher heterozygous mutation (GA genotype), indicating that p.S12N might be a significant ABPA-susceptibility locus ( aspergillus sensitized asthma: OR: 3.02, P = 0.009; aspergillus unsensitized asthma: OR: 2.94, P = 0.005). The mutant allele was preferentially expressed in ABPA patients with heterozygous CARD9S12N , which contributes to its functional alterations to facilitate Af -induced T H 2-mediated ABPA development. In terms of mechanism, Card9 wild-type ( Card9WT ) expression levels decreased significantly due to Af -induced decay of its messenger RNA compared to the heterozygous Card9S12N . In addition, ABPA patients with heterozygous CARD9S12N had increased Af -induced interleukin-5 production. CONCLUSION Our study provides the genetic evidence showing that the heterozygous mutation of CARD9S12N , followed by allele expression imbalance of CARD9S12N , facilitates the development of ABPA.
Humans ; Aspergillosis, Allergic Bronchopulmonary/complications* ; Aspergillus fumigatus/genetics* ; Asthma/genetics* ; Aspergillus ; Mutation/genetics* ; CARD Signaling Adaptor Proteins/genetics*

Objective:To analyze the molecular epidemiological characteristics of the Corona virus disease 2019 (COVID-19) cases in Shijiazhuang, which can reveal the origin of the outbreak and provide a scientific basis for COVID-19 prevention and control.Methods:From January 2 to January 8, 2021, a total of 404 samples from 170 COVID-19 cases were collected from the Shijiazhuang Fifth Hospital. The consensus sequence of 2019 novel Coronavirus(2019-nCoV) was obtained through multiplex polymerase chain reaction-based sequencing. The sequences of 170 COVID-19 cases were analyzed by the PANGOLIN, and the data were statistically analyzed by T-test.Results:Among the 404 COVID-19 samples, a total of 356 samples obtained high quality genome sequences (>95%,100×sequencing depth). The whole genome sequences of 170 COVID-19 cases were obtained by eliminating repeated samples. All 170 sequences were recognized as lineage B1.1 using PANGOLIN. The number of single nucleotide polymorphism arrange from 18-22 and most of the single nucleotide polymorphism were synonymous variants. All of 170 genomes could be classified into 48 sub-groups and most of the genomes were classified into 2 sub-groups (66 and 31, respectively).Conclusions:All cases in this study are likely originated from one imported case. The viruses have spread in the community for a long time and have mutated during the community transmission.

The importance of structural variants(SVs)for human phenotypes and diseases is now recognized.Although a variety of SV detection platforms and strategies that vary in sensitivity and specificity have been developed,few benchmarking procedures are available to confidently assess their performances in biological and clinical research.To facilitate the validation and application of these SV detection approaches,we established an Asian reference material by characterizing the genome of an Epstein-Barr virus(EBV)-immortalized B lymphocyte line along with identified benchmark regions and high-confidence SV calls.We established a high-confidence SV callset with 8938 SVs by integrating four alignment-based SV callers,including 109x Pacific Biosciences(PacBio)continuous long reads(CLRs),22 x PacBio circular consensus sequencing(CCS)reads,104x Oxford Nanopore Technologies(ONT)long reads,and 114×Bionano optical mapping plat-form,and one de novo assembly-based SV caller using CCS reads.A total of 544 randomly selected SVs were validated by PCR amplification and Sanger sequencing,demonstrating the robustness of our SV calls.Combining trio-binning-based haplotype assemblies,we established an SV benchmark for identifying false negatives and false positives by constructing the continuous high-confidence regions(CHCRs),which covered 1.46 gigabase pairs(Gb)and 6882 SVs supported by at least one diploid haplotype assembly.Establishing high-confidence SV calls for a benchmark sample that has been characterized by multiple technologies provides a valuable resource for investigating SVs in human biology,disease,and clinical research.

Objective:To assess the predictive value of histogram parameters derived from synthetic MRI for extramural venous invasion (EMVI) of rectal cancer.Methods:Totally 76 patients with pathologically proven rectal adenocarcinoma were enrolled in this retrospective study from November 2018 to December 2019 in Cancer Hospital, Chinese Academy of Medical Sciences. All participants underwent preoperative rectal MRI examination including MAGiC within 4 weeks before surgery. The status of EMVI on MRI was independently assessed by one junior and one senior radiologist. Histogram parameters were extracted from T 1, T 2 and proton density (PD) mapping, including mean, variance, maximum, minimum, 10 th percentile, median, 90 th percentile, energy, kurtosis, entropy and skewness. With postoperative pathological result as the gold standard, the patients were divided into EMVI-positive group ( n=18) and EMVI-negative group ( n=58). The Mann-whitney U test was used to compare the differences in histogram parameters between the two groups. The ROC curves were used to explore the predictive performance for assessing EMVI. The logistic regression analysis was used to combine the assessment of radiologists with parameters whose area under the ROC curve (AUC)>0.7. The Delong test was used to analyze the differences of diagnostic efficacy between different methods in predicting EMVI. Results:Significant differences of the energy of T 1, T 2 and PD mapping and skewness of PD mapping were observed between the EMVI-positive and EMVI-negative group ( P<0.05), with the AUC of 0.744, 0.728, 0.708 and 0.652. The AUC of junior radiologist in evaluating EMVI was 0.711, and the AUC of the combination with energy of T 1, T 2 and PD mapping was 0.817, showing a statistically significant difference ( Z=2.281, P=0.023). The AUC of senior radiologist in evaluating EMVI was 0.837, and the AUC of the combination with energy was 0.856. There was a significant difference in AUC between junior and senior radiologists in assessing EMVI ( Z=2.587, P=0.010), while there was no significant difference between junior radiologist combined with energy and senior radiologist ( Z=0.578, P=0.563). Conclusion:The histogram parameters based on quantitative mapping of synthetic MRI were useful for predicting EMVI of rectal cancer.

Objective:To assess the optimal cut-off value between early recurrence and late recurrence of patients with hepatocellular carcinoma (HCC) after radiofrequency ablation (RFA), and to construct a nomogram to predict early recurrence.Methods:A total of 119 patients with HCC who recurred after RFA in Cancer Hospital, Chinese Academy of Medical Sciences from January 2012 to December 2017 were identified. The optimal cut-off value to distinguish early and late recurrence was determined based on differences in post recurrence survival (PRS) by minimum P-value approach. The clinical and radiographic risk factors for early recurrence were identified by univariate and multivariate Logistic regression analysis. The predictive nomogram was constructed by these factors and internally validated. Results:The optimal cut-off value to distinguish early recurrence and late recurrence was 12 months after RFA ( P=0.005). The patients were divided into early recurrence group (47 cases) and late recurrence group (72 cases). The lower quartile PRS (Q1-PRS) and lower quartile overall survival (Q1-OS) were 11.1 and 19.1 months in the early recurrence group, which were shorter than 31.6 and 81.0 months in the late recurrence group ( P=0.005 and P<0.001, respectively). The independent risk factors of early recurrence were alpha fetoprotein (AFP) ( OR=8.459, 95% CI: 2.231-32.073), albumin(ALB) ( OR=0.251, 95% CI: 0.047-1.339), number of lesions ( OR=3.842, 95% CI: 1.424-10.365) and peritumoral enhancement ( OR=8.05, 95% CI: 1.23-52.80), which were further incorporated into constructing the predictive nomogram of early recurrence of HCC after RFA. Internal validation results showed the area under the curve, sensitivity, specificity of the receiver operating characteristic (ROC) curve were 0.839, 68.1% and 93.1%, respectively. The calibration curve showed the predicted curve of nomogram was close to the ideal curve. Hosmer-Lemeshow test showed there was no significant difference between the predicted results of nomogram and the actual results ( P=0.424). Conclusions:An interval of 12 months after RFA is the optimal cut-off value for defining early recurrence and late recurrence. The nomogram is integrated by clinical and radiographic features, which can potentially predict early recurrence of HCC after RFA and may offer useful guidance for individual treatment or follow up.

Objective:To assess the optimal cut-off value between early recurrence and late recurrence of patients with hepatocellular carcinoma (HCC) after radiofrequency ablation (RFA), and to construct a nomogram to predict early recurrence.Methods:A total of 119 patients with HCC who recurred after RFA in Cancer Hospital, Chinese Academy of Medical Sciences from January 2012 to December 2017 were identified. The optimal cut-off value to distinguish early and late recurrence was determined based on differences in post recurrence survival (PRS) by minimum P-value approach. The clinical and radiographic risk factors for early recurrence were identified by univariate and multivariate Logistic regression analysis. The predictive nomogram was constructed by these factors and internally validated. Results:The optimal cut-off value to distinguish early recurrence and late recurrence was 12 months after RFA ( P=0.005). The patients were divided into early recurrence group (47 cases) and late recurrence group (72 cases). The lower quartile PRS (Q1-PRS) and lower quartile overall survival (Q1-OS) were 11.1 and 19.1 months in the early recurrence group, which were shorter than 31.6 and 81.0 months in the late recurrence group ( P=0.005 and P<0.001, respectively). The independent risk factors of early recurrence were alpha fetoprotein (AFP) ( OR=8.459, 95% CI: 2.231-32.073), albumin(ALB) ( OR=0.251, 95% CI: 0.047-1.339), number of lesions ( OR=3.842, 95% CI: 1.424-10.365) and peritumoral enhancement ( OR=8.05, 95% CI: 1.23-52.80), which were further incorporated into constructing the predictive nomogram of early recurrence of HCC after RFA. Internal validation results showed the area under the curve, sensitivity, specificity of the receiver operating characteristic (ROC) curve were 0.839, 68.1% and 93.1%, respectively. The calibration curve showed the predicted curve of nomogram was close to the ideal curve. Hosmer-Lemeshow test showed there was no significant difference between the predicted results of nomogram and the actual results ( P=0.424). Conclusions:An interval of 12 months after RFA is the optimal cut-off value for defining early recurrence and late recurrence. The nomogram is integrated by clinical and radiographic features, which can potentially predict early recurrence of HCC after RFA and may offer useful guidance for individual treatment or follow up.

Objective The aim of this study was to determine whether multi?parameters MRI of tongue carcinoma have the potential to predict cervical lymph node metastases. Methods A total of 46 patients with tongue carcinoma, who underwent MRI scan preoperatively, were investigated retrospectively and were divided into cervical lymph node (LN) metastases group (unilateral LN+, n=16;bilateral LN+, n=14) and no cervical lymph node metastases group (LN-, n=16) according to their pathological grading. Of the 40 patients with tongue carcinoma underwent plain and contrast MRI scan, 6 patients have plain MRI scan, and 32 have DWI examination.The ADC value, tumor length, tumor thickness, sublingual distance between tumor and sublingual space, and para?lingual distance between tumor and tongue midlinedetermined from MRI, were preoperatively estimated and compared with the pathological findings of cervical lymph nodes. A unpaired t test was used to analyze normal distributed continuous data, and a Mann?Whitney U test was used to analyze abnormally distributeddata. The ROC was used to evaluate the efficacy of MRI in predicting the metastasis of cervical lymph nodes. Results The indexes of ADC value, tumor length, tumor thickness, and para?lingual distance between tumor and tongue midline, which all showed significant difference between LN+group and LN-group (all P<0.05), and the index of sublingual distance between tumor and sublingual space showed no significantly association with LN+ (P>0.05). The index of ADC value showed significant difference between unilateral LN+group and bilateral LN+group (P<0.05), and the other indexes, which all showed no significantly association with bilateral LN+ (all P>0.05). The ROC curve analysis of the ADC value, tumor length, tumor thickness, and para?lingual distance between tumor and tongue midline of the neck lymph node metastasis were carried out, with the cutoff set as 1.13×10?3 mm2/s, 31.08 mm, 17.33 mm and-2.26 mm. The corresponding area under curve(AUC), sensitivity, and specificity were 0.878, 90.9%and 90.0%; 0.822, 83.3% and 81.3%; 0.834, 86.7% and 81.3%; 0.794, 86.7% and 75.0%, respectively. The ROC curve analysis of the ADC of the bilateral neck lymph node metastasis was also carried out, with the cutoff of ADC value set as 1.07×10?3 mm2/s, the corresponding AUC, sensitivity, and specificity were 0.806, 80.0%and 75.0%. Conclusion The ADC value, tumor length ,tumor thickness and para?lingual distance between tumor and tongue midline,determined from MR imaging, all can be used as independent factors in predicting cervical lymph node metastasis, where ADC value may be helpful to predict bilateral neck lymph node metastasis.

O bjective To investigate the value of diffusion weighted imaging (DWI) in predicting the efficacy of preoperative chemoradiotherapy (CRT) for locally advanced rectal cancer.Methods From 2007 to 2012,86 patients with histopathologicaIly proven rectal cancer who underent pre-CRT were enrolled in this study prospectively.Diffusion-weighted MRl was performed in all patients before pre-CRT, while it was performed in part of the patients during and after pre-CRT as well.ADC values of the tumors were calculated on the workstation.Patients were assigned to the tumor downstaged group or the tumor nondownstaged group on the basis of T staging.The change in ADC following treatment and the difference in ADC between groups were analyzed.Results Of the 86 patients after surgery, 20 were diagnosed with T0, 2 with T1, 17 with T2, 44 with T3 and 3 with T4.39 patients were classified as the downstaged group, of which 18 were of pCR.The remaining 47 patients were classified as the nondownstaged group.Of the total of 86 patients, the mean ADC values before, during, and after pre-CRT (pre-ADC, during-ADC, and post-ADC) were (1.03± 0.17)×10-3 , (1.39±0.28) ×10-3 , and (1.61±0.27) ×10-3 mm 2 /s and there was a significant difference (P<0.001).However, the pre-CRT ADC of the downstaged group did not differ significantly from that of the nondownstaged group (P=0.615).Of the 43 patients who underwent MRI before, during and after pre-CRT, the mean ADC values were (1.05±0.16) ×10-3 , (1.39±0.29) ×10-3 and (1.67±0.30) ×10-3 mm 2 /s, respectively, showing a significant difference (P<0.001) as well.Conclusions The mean ADC value of rectal cancer is gradually increasing along with the course of chemotherapy.Pre-ADC is not a good parameter to be used to predict the efficacy of pre-CRT for locally advanced rectal cancer.

O bjective To investigate the value of diffusion weighted imaging (DWI) in predicting the efficacy of preoperative chemoradiotherapy (CRT) for locally advanced rectal cancer.Methods From 2007 to 2012,86 patients with histopathologicaIly proven rectal cancer who underent pre-CRT were enrolled in this study prospectively.Diffusion-weighted MRl was performed in all patients before pre-CRT, while it was performed in part of the patients during and after pre-CRT as well.ADC values of the tumors were calculated on the workstation.Patients were assigned to the tumor downstaged group or the tumor nondownstaged group on the basis of T staging.The change in ADC following treatment and the difference in ADC between groups were analyzed.Results Of the 86 patients after surgery, 20 were diagnosed with T0, 2 with T1, 17 with T2, 44 with T3 and 3 with T4.39 patients were classified as the downstaged group, of which 18 were of pCR.The remaining 47 patients were classified as the nondownstaged group.Of the total of 86 patients, the mean ADC values before, during, and after pre-CRT (pre-ADC, during-ADC, and post-ADC) were (1.03± 0.17)×10-3 , (1.39±0.28) ×10-3 , and (1.61±0.27) ×10-3 mm 2 /s and there was a significant difference (P<0.001).However, the pre-CRT ADC of the downstaged group did not differ significantly from that of the nondownstaged group (P=0.615).Of the 43 patients who underwent MRI before, during and after pre-CRT, the mean ADC values were (1.05±0.16) ×10-3 , (1.39±0.29) ×10-3 and (1.67±0.30) ×10-3 mm 2 /s, respectively, showing a significant difference (P<0.001) as well.Conclusions The mean ADC value of rectal cancer is gradually increasing along with the course of chemotherapy.Pre-ADC is not a good parameter to be used to predict the efficacy of pre-CRT for locally advanced rectal cancer.