ObjectiveTo investigate the alleviating effect of calcium cyanamide （CaCN₂） soil fumigation on replanting problems of Rehmannia glutinosa. MethodsNewly soil （NP） was used as the control group， while three treatment groups were established： replanted soil （RP）， newly soil treated with CaCN₂ （120 g·m²， tillage depth 25 cm） （NPCC）， and replanted soil treated with CaCN₂ （RPCC）. R. glutinosa was cultivated in all groups. At harvest， the tuber agronomic traits （number of enlarged roots， maximum root diameter， fresh weight， dry weight） were measured. The content of catalpol and rehmannioside D was quantified by ultra-high-performance liquid chromatography （UPLC） to evaluate medicinal quality. Rhizosphere soil available nutrients and enzyme activities were analyzed by assay kits. The community structure and composition of fungi and bacteria in rhizosphere soil were assessed via internal transcribed spacer 2 （ITS2） sequencing and 16S rDNA sequencing， respectively. ResultsCompared with NP， the RP group showed obviously reduced in tuber agronomic traits and quality indicators （P0.05）. However， the RPCC group showed significant improvement in agronomic traits and a notable increase in rehmannioside D content compared to RP （P0.05）. The contents of available phosphorus and potassium in RPCC and NP groups were obviously lower than those in RP （P0.05）. The polyphenol oxidase soil （S-PPO） activity in RP was obviously lower than in NP （P0.05）， while sucrose soil （S-SC）， acid phosphatase soil （S-ACP）， and S-PPO activities in RPCC were obviously higher than in RP （P0.05）. Microbial richness and diversity in RP were obviously higher than in NP （P0.05）， whereas no significant differences were observed between the RPCC and NP. The relative abundances of fungal genera Nectria， Myrothecium， Tomentella， and bacterial genus Skermanella were obviousl lower in RPCC and NP than in RP （P0.05）. Correlation analysis that S-ACP activity was positively correlated with the content of rehmannioside D （P0.05）. Fungal genera Engyodontium and Alternaria， and bacterial genera Pir4 lineage， Pirellula， Methyloversatilis， Brevundimonas， Ralstonia， and Acidibacter were obviously positively correlated with tuber dry weight （P0.05）. Conversely， fungal genera Pseudaleuria， Nectria， Haematonectria， Ceratobasidium， and bacterial genera Streptomyces， Skermanella， RB41， Gemmatimonas， and Bacillus were obviously negatively correlated with dry weight （P0.05）. The fungal genus Alternaria and bacterial genera Brevundimonas， Ralstonia， Acidibacter， and Dongia showed positive correlations with medicinal quality of R.glutinosa tuber， while fungal genera Pseudaleuria， Nectria， Stachybotrys， Fusarium， Gibberella， Ceratobasidium， and bacterial genera Sphingomonas， Skermanella， RB41， Gemmatimonas， and Bacillus were obviously negatively correlated （P0.05）. ConclusionCaCN₂ soil fumigation can significantly improve enzyme activities in replanted Rehmannia rhizosphere soil， enhance the utilization of available nutrients， reshape microbial community structure of replanted R.glutinosa at the family and genus level， and notably improve tuber agronomic traits and medicinal quality. This study provides a novel approach to alleviating replanting problems and offers insights for the integrated development of standardized cultivation techniques， including soil disinfection， nutrient-targeted regulation， and microbial inoculant application.

ObjectiveTo establish a rapid method for evaluating the heterozygosity of Murraya paniculata germplasm materials and provide as a foundation for developing germplasm breeding and innovation measures for M. paniculata. MethodSingle nucleotide polymorphisms （SNPs） were screened from the genome resequencing data of 65 plants of M. paniculata. A self-written script was used to transform 20 SNPs into restriction fragment length polymorphism （RFLP） markers. Polymerase chain reaction-restriction fragment length polymorphism （PCR-RFLP） was employed to detect the 20 RFLP markers in 12 M. paniculata germplasm accessions， and the heterozygosity of M. paniculata germplasm accessions was calculated based on the number of enzyme-cutting bands at the 20 RFLP marker sites. Plink was used to calculate the whole genome heterozygosity of 12 M. paniculata germplasm accessions， and the results obtained with different methods were compared. ResultThere was no significant difference in the heterozygosity calculated by the PCR-RFLP method and the genome resequencing method. The PCR-RFLP and genome resequencing methods identified 8 and 9 germplasm accessions， respectively， with a heterozygosity level less than 30%. Seven germplasm accessions with heterozygosity less than 30.00% were calculated by both methods. ConclusionThe PCR-RFLP method established in this study for evaluating the heterozygosity of M. paniculata germplasm demonstrates the precision of 87.5% and the accuracy of 77.8%. This method serves as a reference for developing heterozygosity evaluation methods in other medicinal plant germplasm resources.

Objective To investigate the effect and underlying mechanisms of G-protein-coupled receptor 39(GPR39)activation on neuroinflammation and brain injury after experimental intracerebral hemorrhage in mice.Methods Mouse model of intracerebral hemorrhage(ICH)was established by intracerebral injection of autologous blood.A total of 176 male C57/BL6 mice were randomly divided into 8 groups:Sham group(n=42),ICH group(n=34),ICH+Vehicle group(n=32),ICH+TC-G 1008 group(n=44),ICH+GPR39 siRNA group(n=6),ICH+Scramble siRNA group(n=6),ICH+TC-G 1008+666-15 group(n=6),and ICH+TC-G 1008+Vehicle 2 group(n=6).GPR39-specific agonist TC-G 1008 was administered via oral gavage at 1 and 25 h post-ICH modeling.Additionally,GPR39 siRNA and cAMP response element binding protein(CREB)inhibitor 666-15 were intracerebroventricularly injected 24 h before induction of ICH to inhibit the expression levels of GPR39 and p-CREB.At 48 h after ICH,modified Garcia test,forelimb placement test and corner turn test were used to evaluate the short-term neurological deficits in mice.Brain water content was determined by wet/dry method.Immunofluorescence assay was performed to detect the co-localization of GPR39 in neurons and microglia in the brain tissue surrounding the hematoma,as well as the expression of NOD-like receptor thermal protein domain associated protein 3(NLRP3)in neurons.ELISA was employed to measure IL-1β,TNF-α and myeloperoxidase(MPO)levels in peri-hematoma tissue.TUNEL staining was performed to quantify apoptotic neurons around the hematoma.Nissl staining was utilized to evaluate neuronal damage.Western blotting was conducted to detect the expression of GPR39,p-CREB,CREB,NLRP3,Cleaved caspase-1(C-caspase-1),and gasdermin-D protein(GSDMD)in peri-hematoma brain tissue.Results GPR39 expression peaked at 48 h post-ICH in mice(P＜0.05),and it was expressed in both neurons and microglia.Activation of GPR39 by TC-G 1008(24 mg/kg)significantly improved the modified Garcia score,and increased success rate of left forelimb placement and the number of left turns(P＜0.05).Brain edema in the ipsilateral basal ganglia(BG)and cortex(CX)was significantly reduced(P＜0.05).The numbers of apoptotic and damaged neurons around the hematoma were obviously decreased(P＜0.05).The expression of pyroptosis-related molecules,including NLRP3,C-caspase-1 and GSDMD and the levels of inflammation-related factors,including IL-1β,TNF-α and MPO were notably decreased(P＜0.05).However,knockdown of GPR39 and downregulation of p-CREB expression significantly increased the expression of pyroptosis related molecules and inflammatory-related factors in peri-hematoma brain tissue post-ICH in mice(P＜0.05).Conclusion GPR39 activation may inhibit neuroinflammation and brain injury after ICH in mice partly through the CREB signaling pathway.Therefore,GPR39 may be a potential therapeutic target for mitigating neuroinflammation and brain damage after ICH.

Rheumatic mitral stenosis is one of the most common cardiac valvulopathies in our country, which is relatively rare in European and American countries. Medical therapy is reserved mainly for the treatment of complications, which can not fundamentally change the valve structure. Only surgical treatments can correct these valve lesions, including closed mitral commissurotomy, percutaneous balloon mitral valvuloplasty, mitral valve repair under direct vision and mitral valve replacement. Numerous studies demonstrate that valve repair provides better long-term results, though it occupies a low proportion clinically. This article reviewed domestic and foreign literature concerning surgical treatments for patients with rheumatic mitral stenosis to provide some reference for the peers.

Objective:To evaluate the value of radiomics based on contrast-enhanced spectral mammography (CESM) of internal and peripheral regions combined with clinical factors in predicting benign and malignant breast lesions of breast imaging reporting and data system category 4 (BI-RADS 4).Methods:A retrospective analysis was performed on the clinical and imaging data of patients with breast lesions who were treated in Yantai Yuhuangding Hospital (Center 1) Affiliated to Qingdao University from July 2017 to July 2020 and in Fudan University Cancer Hospital (Center 2) from June 2019 to July 2020. Center 1 included 835 patients, all female, aged 17-80 (49±12) years, divided into training set (667 cases) and test set (168 cases) according to the "train-test-split" function in Python software at a ratio of 8∶2; and 49 patients were included from Center 2 as external validation set, all female, aged 34-70 (51±8) years. The radiomics features were extracted from the intralesional region (ITR), the perilesional regions of 5, 10 mm (PTR 5 mm, PTR10 mm) and the intra-and perilesional regions of 5, 10 mm (IPTR 5 mm, IPTR 10 mm) and were selected by variance filtering, SelectKBest algorithm, and least absolute shrinkage and selection operator. Then five radiomics signatures were constructed including ITR signature, PTR 5 mm signature, PTR 10 mm signature, IPTR 5 mm signature, IPTR 10 mm signature. In the training set, univariable and multivariable logistic regressions were used to construct nomograms by selecting radiomics signatures and clinical factors with significant difference between benign and malignant BI-RADS type 4 breast lesions. The efficacy of nomogram in predicting benign and malignant BI-RADS 4 breast lesions was evaluated by the receiver operating characteristic curve and area under the curve (AUC). Decision curve and calibration curve were used to evaluate the net benefit and calibration capability of the nomogram.Results:The nomogram included ITR signature, PTR 5 mm signature, PTR 10 mm signature, IPTR 5 mm signature, age, and BI-RADS category 4 subclassification for differentiating malignant and benign BI-RADS category 4 breast lesions and obtained AUCs of 0.94, 0.92, and 0.95 in the training set, test set, and external validation set, respectively. The calibration curve showed good agreement between the predicted probabilities and actual results and the decision curve indicated a good net benefit of the nomogram for predicting malignant BI-RADS 4 lesions in the training set, test set, and external validation set.Conclusion:The nomogram constructed from the radiomics features of the internal and surrounding regions of CESM breast lesions combined with clinical factors is attributed to differentiate benign from malignant BI-RADS category 4 breast lesions.

ObjectiveTo establish a rapid screening method for germplasm materials of Gastrodia elata with high purity， and lay a foundation for pure line breeding and cross breeding. MethodBased on the whole genome sequencing and population resequencing of G. elata， 20 restriction fragment length polymorphism (RFLP) markers were developed by single nucleotide polymorphism (SNP) sites. The polymerase chain reaction (PCR)-RFLP method was used to carry out restriction endonuclease experiments on 20 RFLP markers of 15 G. elata germplasms. According to the number of enzymatic bands at 20 RFLP marker sites, the purity of 15 germplasms was calculated and evaluated. On this basis, genome resequencing technology was used to verify the assessment results. ResultTen germplasm materials with purity greater than 95% were screened out by PCR-RFLP method， 3 of which had 95% purity and 7 had 100% purity. Nine germplasm materials with purity greater than 95% were screened out by genome resequencing methods, and 8 of them were consistent with the results of PCR-RFLP. ConclusionThe PCR-RFLP method established in this study for screening G. elata germplasms with high purity precision of RFLP markers has 80% precision and 89% accuracy. The method is simple， efficient， and significantly less expensive than genome resequencing method, which provides technical support for pure line breeding of G. elata and references for breeding of other Chinese medicinal materials.

ObjectiveTo analyze the changes of volatile organic compounds （VOCs） and bacterial community characteristics in rhizosphere soil during the growth of Rehmanniae Radix， as well as the relationship between VOCs and bacterial community structure， so as to lay the foundation for the evaluation of the characteristics of continuous cropping soil and the regulation of continuous cropping soil microorganisms. MethodThe rhizosphere soil during the three main growth periods of Rehmanniae Radix in July， August and October was used as the research object. Gas chromatography-mass spectrometry （GC-MS） was used to determine the relative contents of VOCs in ethyl acetate and dichloromethane parts in rhizosphere soil. The characteristics of rhizosphere bacterial community structure was determined by high-throughput sequencing， SPSS 24， SIMCA 14.1 and other software were used to analyze the differences of VOCs and bacterial community structure in rhizosphere soil of the three periods， and the main VOCs and iconic bacteria that caused the differences were screened， and the correlation between VOCs and bacterial community structure was analyzed. ResultThere were differences in VOCs in different parts of rhizosphere soil during the three growth stages of Rehmanniae Radix. Among them， and the main VOCs that cause differences were dioctyl isophthalate， 2-octyl-1-dodecanol and 2-ethylhexyl p-toluic acid in the ethyl acetate part， and 1，3-benzenedicarboxylic acid， di （2-propylpentyl） phthalate and 2-ethylhexyl p-toluenecarboxylic acid in the dichloromethane part. From the seedling stage to the end of tuber enlargement of Rehmanniae Radix， the relative abundance of dominant bacteria such as Actinobacteria， Firmicutes and Chloroflexi in the rhizosphere soil was gradually decreased， and there were unique bacterial communities in rhizosphere soil of each growth stage. Correlation analysis showed that the VOCs in rhizosphere soil of Rehmanniae Radix had an impact on the rhizosphere bacterial community structure， especially the components of esters and alcohols. ConclusionDuring the growth of Rehmanniae Radix， the characteristics of rhizosphere soil are mainly manifested in the content changes of main VOCs such as esters and alcohols and the gradual decrease of the abundance of the main beneficial bacteria， and the VOCs in rhizosphere soil play a certain role in shaping the structure of bacterial community.

Plants with alien genomic components (alien chromosomes / chromosomal fragments / genes) are important materials for genomic research and crop improvement. To date, four strategies based on trait observation, chromosome analysis, specific proteins, and DNA sequences have been developed for the identification of alien genomic components. Among them, DNA sequence-based molecular markers are mainly used to identify alien genomic components. This review summarized several molecular markers for identification of alien genomic components in wheat, cabbage and other important crops. We also compared the characteristics of nine common molecular markers, such as simple sequence repeat (SSR), insertion-deletion (InDel) and single nucleotide polymorphism (SNP). In general, the accuracy of using a combination of different identification methods is higher than using a single identification method. We analyzed the application of different combination of identification methods, and provided the best combination for wheat, brassica and other crops. High-throughput detection can be easily achieved by using the new generation molecular markers such as InDel and SNP, which can be used to determine the precise localization of alien introgression genes. To increase the identification efficiency, other new identification methods, such as microarray comparative genomic hybridization (array-CGH) and suppression subtractive hybridization (SSH), may also be included.
Chromosomes, Plant ; Comparative Genomic Hybridization ; Genome, Plant/genetics* ; Genomics ; Triticum/genetics*

Objective:To evaluate the value of pretreatment 18F-fluorodeoxyglucose (FDG) PET/CT-based heterogeneity for early prediction of targeted therapy outcome in patients with human epidermal growth factor receptor 2 (HER2) positive metastatic breast cancer. Methods:From May 2012 to April 2018, 29 patients (all females, median age: 52 (32-69) years) who had HER2 positive metastatic breast cancer and underwent pretreatment 18F-FDG PET/CT in Fudan University Shanghai Cancer Center were retrospectively enrolled. All patients received trastuzumab as first-line treatment and were followed up for 6-87 (median time: 35) months. The relations between clinicopathologic parameters or PET/CT-based parameters and progression-free survival (PFS)/overall survival (OS) were analyzed with Cox univariate analysis. The parameters with P≤0.01 were further analyzed with Cox multivariate analysis. Optimal cut-off values were determined by time-dependent receiver operating characteristic (ROC) curve analysis. The survival analyses were estimated by Kaplan-Meier method and log-rank test. Results:The median OS of the 29 patients was 30 (6-83) months, and the median PFS was 10 (2-29) months. The PET/CT-based heterogeneity index(HI), including the maximum standardized uptake value (SUV max) ratio (SUV max-R; hazard ratio ( HR)=8.6, 95% CI: 2.7-27.8, P<0.001), the mean standardized uptake value (SUV mean)-2.5 (the cut-off value of standardized uptake value (SUV)=2.5) ratio (SUV mean-2.5-R; HR=2.6, 95% CI: 1.2-5.9, P=0.020), the metabolic tumor volume(MTV)-2.5 ratio(MTV-2.5-R; HR=2.4, 95% CI: 1.1-5.2, P=0.030), and the total lesion glycolysis(TLG)-2.5 ratio(TLG-2.5-R; HR=3.2, 95% CI: 1.4-7.4, P=0.008) of the lesion with the highest SUV max to that with the lowest SUV max, were significantly associated with PFS. None of the parameters was significantly associated with OS (all P>0.05). Multivariate analysis showed that the SUV max-R was the only independent predictor for PFS ( HR=6.8, 95% CI: 1.8-26.1, P<0.01). Area under the ROC curve for SUV max-R was 0.747. With a cut-off value of 1.8, SUV max-R could effectively distinguish the benefit from non-benefit population treated with trastuzumab (15.0 vs 7.0 months; χ2=18.68, P<0.01). Conclusion:Pretreatment 18F-FDG PET/CT-based HI has potential value for early prediction of first-line trastuzumab treatment outcome in patients with HER2 positive metastatic breast cancer.

Objective:To investigate the diagnostic performance of repeat biopsy 68Ga-PSMA PET/CT to distinguish between benign and malignant prostate disease. Methods:The clinical data and medical imaging of thirty-nine patients underwent repeat prostate biopsy were analyzed respectively in this study. The median age of patients was 65 years (range 46-81 years), the median PSA level was 11.0ng/ml (range 5.4-49.8 ng/ml), f/tPSA was 0.15(0.01-16.50)ng/ml, prostate volume was 43.80(7.79-108.63)ml, and PSA density was 0.24(0.09-2.31)ng/ml 2. All patients underwent pre-biopsy 68Ga-PSMA PET/CT and the standard transrectal ultrasound-guided systematic prostate biopsy. Based on the biopsy results, 68Ga-PSMA PET/CT images of all patients were visually and semi-quantitatively analyzed. By visual analysis, 68Ga-PSMA uptake in prostate was defined as focal, multimodal and inhomogeneous, and then the detection rate of prostate cancer in each subgroup was analyzed. The value of the ROC curve in the diagnosis of prostate cancer was analyzed based on the SUV max of prostate cancer(SUV max), tumor-to-normal-prostate background(SUV T/BGp)as semi-quantitative parameters of 68Ga-PSMA PET/CT. Results:Prostate cancer was detected in 18 patients (46.2%) and 12 patients (30.8%) had clinically significant disease. There were 11, 5 and 2 patients with prostate cancer respectively in men with a focal (12 patients), multifocal(7 patients) and inhomogeneous (20 patients) 68Ga-PSMA uptake. The ROC analysis revealed a SUV max 5.3 and SUV T/BGp1.8 as an optimal cut-off level to distinguish between non-prostate cancer and prostate cancer in 68Ga-PSMA PET/CT, the sensitivity and specificity were 100.0% and 85.7% for SUV max (AUC=0.979), 83.3% and 90.5% for SUV T/BGp (AUC=0.915). Conclusions:Pre-biopsy 68Ga-PSMA PET/CT could help to distinguish between benign and malignant prostate disease before repeat prostate biopsy and detect the foci of prostate cancer.