Pinellia ternata， belonging to the Pinellia genus within the Araceae family， is a medicinal plant due to its tubers. There are severe issues with unclear germplasm and mixed varieties in its cultivation， necessitating urgent new variety protection efforts. The distinctness， uniformity， and stability （DUS） testing of the plant variety is the basis for protecting new plant varieties， and the DUS test guidelines are the technical basis for DUS testing. To develop the DUS test guidelines for P. ternata， agronomic traits of 229 germplasm of P. ternata were observed and measured during its two growth stages over the years， and each character was graded and described. A total of 38 traits were selected as the test traits of the DUS test guideline for P. ternata. There were three plant traits， 19 leaf traits， six flower traits， two fruit traits， two tuber traits， five bulbil traits， and one ploidy trait. These traits could be divided into 22 quality characters， 12 quantitative characters， and four pseudo-quantitative characters， as well as seven groups， including plants， leaves， flowers， fruit， tubers， bulbils， and ploidy. By searching for standard traits， 10 standard varieties were ultimately determined. Preparing these guidelines will have great significance for reviewing and protecting P. ternata varieties， safeguarding breeders' rights， and promoting the development of the P. ternata industry.

ObjectiveTo explore the genetic relationship and reveal the genetic variations of 45 germplasm accessions of Artemisia argyi. MethodGenotyping-by-sequencing （GBS） was employed to mine single nucleotide-polymorphisms （SNPs） from the 45 germplasm accessions. Principal component analysis， phylogenetic analysis， population genetic structure analysis， and genetic variation analysis were conducted based on the SNPs. ResultA total of 111.91 Gb of data were obtained， with the Q20， Q30， and average GC content of 96.39%， 90.33%， and 39.37%， respectively. The comparison rate between clean reads and the reference genome was 70.24%-98.97%. A total of 22 399 Indels and 170 539 SNPs were obtained， and the 10^th pair of chromosomes had the most variation sites. The results of principal component analysis， cluster analysis， and genetic diversity analysis classified the 45 germplasm accessions into three groups. Group Ⅰ contained three germplasm accessions from Qichun County. The germplasm accessions in group Ⅱ were all wild. Group Ⅲ contained 31 germplasm accessions， with the most complex sources. Moreover， the 45 germplasm accessions can be classified into 3 subtypes， containing the genetic information from three ancestors. The results indicated rich genetic diversity of A. argyi from different sources， especially the germplasm accessions from Qichun County， Hubei province. ConclusionThis study provides theoretical support for breeding new varieties， developing specific SNP markers， and revealing the genetic relationship of A. argyi.

Objective To explore the mechanism by which Sestrin2(SESN2)regulates autophagy activity of chondrocytes by mediating mammalian rapamycin target protein complex 1(mTORC1)signaling pathway.Methods The normal chondrocytes were treated with interleukin-1 β(IL-1β)to establish an osteoarthritis(OA)chondrocyte model,which was divided into the control group and the IL-1 β-treated group.Real-time quantitative PCR(qPCR)and Western blot were used to detect the expression levels of matrix metalloproteinase 13(MMP13),type Ⅱ collagen(COL2A1)and SESN2 in the two groups.The cell models of the chondrocyte overexpression SESN2 group and knockdown SESN2 group were obtained via cell transfection technology,and the expression levels of SESN2 in each group were detected by qPCR while those of SESN2,MMP13,COL2A1,mTORC1 pathway-related proteins and autophagy-related proteins in each group were detected by Western blot.The effects of SESN2 on cell proliferation and migration were detected by CCK-8 and cell scratch assay.Results(1)The expression level of MMP13 in the IL-1 β-treated group was significantly up-regulated,while the expression levels of COL2A1 and SESN2 were significantly decreased.(2)Compared with the control group,the expressions of p-mTORC1,ribosomal protein S6 kinase 1(S6K1),and MMP13 protein in OA chondrocytes in the overexpression group were significantly down-regulated,while the expressions of adenosine 5'-monophosphate(AMP)-activated protein kinase(AMPK)and chondroprotective gene COL2A1 were significantly increased,and the expression level of Beclin-1 and the ratio of microtubule associated protein 1 light chain 3-Ⅱ(LC3-Ⅱ)/(LC3-Ⅰ)were increased.Meanwhile,overexpression of SESN2 could up-regulate the proliferation and migration of chondrocytes,but the results were opposite after knockdown of SESN2.Conclusion SESN2 can enhance autophagy,proliferation and migration of chondrocytes by inhibiting mTORC1 pathway,which has provided data for revealing the pathogenesis of OA and exploring new therapeutic methods.

Cancer immunotherapy has great potential and is expected to become the mainstream method of cancer treatment.In the current application of cancer immunotherapy,immune checkpoint inhibitors(ICIs)have achieved remarkable results.The cur-rently widely used ICIs in clinical practice include inhibitors targeting cytotoxic T lymphocyte-associated antigen-4(CTLA-4),pro-grammed death-1(PD-1)and programmed death-ligand 1(PD-L1).In addition,new immunotherapies such as oncolytic viruses and chimeric antigen receptor T cells are gradually entering the clinical practice,and combination therapy related to ICIs has shown unique advantages.This article will focus on the current application status of ICIs,oncolytic viruses,and chimeric antigen receptor T cell ther-apies in solid tumors either their individual or combined forms.

【Objective:】 To summarize the research status, problems and countermeasures of ethical issues related to volunteer action in public health emergencies by reviewing relevant literature. 【Methods:】 Using four keywords: public health emergencies, volunteers, ethics, and research, this paper searched the Chinese and English databases for the literature published from December 2006 to December 2021 (15 years in total) on "ethical research on volunteer action in public health emergencies". After searching, it was found that there were not many related papers. A total of 31 papers were retrieved, and only 23 papers were selected. 【Results:】 The ethical problems faced by volunteers in public health emergencies were complex and diverse, and the individual ethical behavior of volunteers was affected by many factors. The strategies to solve the relevant ethical issues were not yet perfect. 【Conclusions:】 The ethical issues related to volunteer actions in public health emergencies needed to be further studied.

Objective:To investigate the relationship between the changes of default network topology properties of brain function and cognitive function in patients with end-stage renal disease (ESRD).Methods:A total of 31 patients with ESRD were enrolled in the Department of Nephrology, Changzhou Second Hospital Affiliated to Nanjing Medical University from January 2019 to December 2020, and 18 healthy persons were included in the same period as the control group.The cognitive function was evaluated with the Montreal cognitive assessment (MoCA) and trail making tests, and then the subjects were examined by resting-state functional magnetic resonance imaging (rs-fMRI). After preprocessing, the brain functional network was constructed and the topology properities of the network were calculated.The SPSS 20.0 software was used for statistical analysis.Independent sample t-test, chi square test and Pearson correlation analysis were used for data statistics. Results:(1) The score of MoCA in the ESRD group(23.37±1.77) was significantly lower than that in the healthy control group(27.94±1.13)( t=9.537, P<0.001). (2) The levels of Eglobal, Elocal, Cp and Sigma in ESRD group ((0.129±0.025), (0.148±0.040), (0.188±0.046), (1.593±0.650)) were significantly lower than those in healthy control group ((0.160±0.040), (0.212±0.024), (0.276±0.049), (2.004±0.864))( t=3.591, 7.474, 7.058, 2.034, all P<0.05). The Lp value of the ESRD group (8.131±1.905) was significantly higher than that of the control group (6.777±2.150)( t=2.583, P< 0.05). The node efficiency values of bilateral dorsolateral superior frontal gyrus, left middle frontal gyrus, bilateral posterior cingulate gyrus, right hippocampus, left superior marginal gyrus, bilateral angular gyrus and bilateral cuneate anterior lobe in ESRD group ((0.133±0.071), (0.201±0.047), (0.211±0.106), (0.175±0.066), (0.276±0.113), (0.122±0.146), (0.042±0.075), (0.171±0.027), (0.154±0.078), (0.240±0.095), (0.161±0.056))were lower than those in the healthy control group((0.312±0.075), (0.289±0.091), (0.277±0.132), (0.284±0.053), (0.368±0.063), (0.231±0.227), (0.120±0.162), (0.296±0.064), (0.310±0.186), (0.318±0.066), (0.286±0.103))( t=2.107-9.436, all P<0.05). (3)Pearson correlation analysis showed that the node efficiency values of bilateral posterior cingulate gyrus and right hippocampus in ESRD group were positively correlated with the score of MoCA( r=0.36, 0.49, 0.53, all P<0.05). Conclusion:The topological structure of brain functional network is abnormal in ESRD patients, which can affect the cognitive function of patients.

ObjectiveTo investigate the effects of different potassium application rates on the growth， yield and quality of two-year-old Belamcanda chinensis and to provide a basis for potassium application to achieve high yield and quality of B. chinensis. MethodOn the basis of nitrogen and phosphorus fertilization at 120 kg·hm^-2， we designed five potassium treatments of 0 （K₀）， 60 （K₆₀）， 120 （K₁₂₀）， 180 （K₁₈₀） and 240 （K₂₄₀） kg·hm^-2 in a field experiment to study the effects of different potassium application rates on the agronomic traits， rhizome yield， flavonoid content and potassium fertilizer utilization efficiency of the two-year-old B. chinensis. ResultThe application of potassium fertilizer promoted the root development and tillering， which significantly improved the yield of B. chinensis. Compared with the K₀ treatment， the other potassium treatments increased the rhizome yield by 16.0%-44.0% and K₁₈₀ had the highest yield. The application of potassium fertilizer increased the accumulation of flavonoids in the rhizomes of B. chinensis. The total accumulation of six flavonoids （mangiferin， tectoridin， iridin， tectorigenin， irigenin and irisflorentin） increased by 33.2% in the K₁₂₀ treatment compared with that in the K₀ treatment. Furthermore， the application of potassium fertilizer promoted the accumulation of N， K， Mg， Fe and Zn. With the increase in potassium application rate， the potassium accumulation of B. chinensis first increased and then decreased， while the partial productivity and the absorptivity of roots and leaves gradually decreased. The agronomic efficiency of potassium fertilizer was higher at low to medium potassium application rates and decreased significantly at high potassium application rates. The results of the principal component analysis indicated that the ranking of the overall quality score of B. chinensis was K₁₂₀>K₁₈₀>K₂₄₀>K₆₀>K₀. ConclusionConsidering factors such as yield， quality and potassium utilization efficiency of two-year-old B. chinensis， the potassium application at 120-166 kg·hm^-2 is recommended for the production of B. chinensis in Tuanfeng County.

ObjectiveTo clarify the accumulation and distribution characteristics of dry matter and mineral elements in Artemisia argyi var. argyi cv. Qiai， and to provide technical support for the high yield of and efficient utilization of nutrients in this medicinal species. MethodTwo cultivars of this species， Qiqing 1 and Qihuang 1 were selected， and the composition of dry matter in different organs， the content， accumulation， and distribution of mineral elements in each organ of the two cultivars， and the dynamic changes of volatile oil content and index components eucalyptol and borneol in leaves of the two cultivars were monitored at different growth stages. ResultThe period from February to March marked the early growth stage of Qiai， and the dry matter was mainly distributed in the leaves. It accelerated the growth in April， and the period from April to mid-June witnessed the vigorous vegetative growth of Qiai， during which the dry matter was mainly found in the stems and leaves. It began the reproductive growth from late June and the dry matter was mainly distributed in the stems. In the flowering stage in August， no dry matter accumulation occurred. As for the volatile oil， the content was high （> 1.10%） at the vigorous vegetative growth stage and peaked on June 14 （1.33% in Qiqing 1， and 1.23% in Qihuang 1）. The relative mass fraction of eucalyptol was the maximum at the vegetative growth stage （8.67% in Qiqing 1， and 13.07% in Qihuang 1）. The relative mass fraction of borneol peaked at the early growth stage （2.63% in Qiqing 1， and 5.94% in Qihuang 1）. The content of nitrogen， phosphorus， potassium， and zinc in leaves was in significantly positive correlation with the content of volatile oil and the relative content of eucalyptol and borneol. The content of macroelements nitrogen， phosphorus， potassium， and calcium and trace elements iron and zinc peaked at the early growth stage， and the content was the highest in stem and leaf. The content of macroelement magnesium and trace elements manganese and copper was the highest at vegetative growth stage when the content of other elements decreased and the nutrients were gradually transferred to the buds， flowers and other organs. In the whole growth period， the distribution of potassium， calcium， and zinc was in the order of leaf > stem > root， and the distribution of nitrogen， phosphorus， copper， magnesium， and manganese followed the order of leaf > root > stem. The distribution of iron was in the order of root > leaf > stem. There was a significantly positive correlation between the total amount of dry matter and the absorption of nutrients in 'Qiai'. The absorption of macroelements by Qiai was in the order of potassium > nitrogen > calcium > phosphorus > magnesium， and the ratio of absorbed elements was about 2.66∶2.51∶0.6∶0.11∶0.04. The absorption of trace elements followed the order of manganese > iron > zinc > copper， and the ratio of absorbed elements was about 0.25∶0.17∶0.05∶0.04. In terms of the production of medicinal materials， 'Qiai' needed about 4.11 kg potassium， 3.58 kg nitrogen， 0.91 kg phosphorus， 0.18 kg calcium， 0.06 kg magnesium， about 6.64 g manganese， 2.56 g iron， 1.30 g zinc， and 0.92 g copper to produce 100 kg medicinal materials. ConclusionEnough organic fertilizer and phosphorus and potassium fertilizers should be applied as base fertilizers for Qiai. The vegetative growth stage （April-June） marks the high accumulation of dry matter and large demand of nutrients， during which topdressing should be conducted timely and early， especially nitrogen fertilizer， and appropriate amount of micro-element fertilizer should be added. Qiai needs a large amount of calcium and magnesium fertilizers from the mid-vegetative growth stage， and they should be applied in time in the late stage to ensure the vegetative growth of the plants for seeds and the quality of the medicinal material of Qiai.

Cytosine methylation is one of the major types of DNA epigenetic modifications and plays an important role in maintaining normal cell function and regulating gene expression. Bisulfite sequencing PCR (BSP) based cloning and sequencing is a general method for detecting DNA methylation at specific sites, which can clarify the methylation status of each CpG site in the target fragment. However, this method requires large amounts of single-clonal sequencing, which is complicated to operate, time consuming and expensive. Therefore, the development of an accurate, efficient and convenient DNA methylation detection technology is of great significance to improve the efficiency of epigenetic research. Based on the high-throughput mutation detection platform Hi-TOM (high-throughput tracking of mutations) developed by our group, we further established a site-specific DNA methylation high-throughput detection platform Hi-Meth (High-throughput Detection of DNA Methylation). After bisulfite treatment of DNA samples, the specific site-specific DNA methylation analysis results could be obtained through the Hi-Meth platform by performing only one round of PCR amplification. Using the Hi-Meth platform, the DNA methylation status of two promoter regions of rice were detected. The DNA methylation results from Hi-Meth were consistent with the results from BSP-based method. Thus, site-specific DNA methylation analysis results could be obtained accurately and conveniently through the Hi-Meth platform. In conclusion, Hi-Meth provides an important methylation detection platform for specific DNA regions, which has important significance for epigenetic research.
DNA Methylation ; Epigenesis, Genetic ; Epigenomics ; High-Throughput Nucleotide Sequencing/methods* ; Polymerase Chain Reaction ; Sequence Analysis, DNA/methods*