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.

Objective To explore the post-traumatic growth course for women with termination of pregnancy due to fetal abnormalities and analyze the promoting factors,in order to provide a basis for nursing and intervention tactics.Methods The purposive sampling and theoretical sampling methods were used to select 16 women who were diagnosed with fetal abnormalities and decided to terminate pregnancy in the obstetrics department of a tertiary care maternity hospital in Beijing from May 2021 to February 2022.Women were interviewed by semi-structured interviews,and the data was analyzed by Charmaz's constructing grounded theory paradigm.Results Women with termination of pregnancy due to fetal abnormalities underwent 4 stages of post-traumatic growth:traumatic stress period,rumination and seeking of help period,and acceptance internalization period,and post-traumatic growth period.The behavior of each period was influenced by the psychological experience of the corresponding period and influenced the experience of the next period.Extroverted personality,previous normal pregnancy and delivery experience,positive perception of events,positive self-identification,good social support,appropriate hospitalization environment and regulations,open mindedness towards death and religious belief,affected women's post-traumatic growth.Conclusion The post-traumatic growth course of women with termination of pregnancy due to fetal anomaly is a complex continuously sublimated process.Medical staff should pay attention to psychological and behavioral changes in women's post-traumatic growth,combine the factors that promote post-traumatic growth,and provide targeted guidance to help them successfully go through this difficult period and achieve personal growth.

Objective:To observe any effect of supplementing applied behavior analysis with speech-and-language therapy in improving the language ability of children on the autism spectrum.Methods:A total of 60 children with an autism spectrum disorder were divided at random into an experimental group ( n=30) and a control group ( n=30). The control group accepted 3 hours of applied behavior analysis 5 days per week for 3 consecutive months. Over the same period the experiment group accepted 1.5 hours of applied behavior analysis and 1.5 hours of speech-language therapy. Before and after the intervention, sign-significate relations (S-S) and the language retardation examination were used to evaluate the language comprehension, expression, oral communication, the complexity of oral expression content, and the vocabulary of comprehension and expression of the two groups. Results:After the intervention, comprehension and expression in each stage of the S-S had improved significantly in both groups, but the average comprehension and language expression of the experimental group was significantly better than that of the control group. Oral communication improved significantly in both groups but complexity and the vocabulary used had improved significantly more in the experimental group.Conclusion:Supplementing applied behavioral analysis with speech-language therapy can improve the language ability of children on the autism spectrum significantly better than behavioral analysis alone.

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.

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*

OBJECTIVETo investigate the effects of long non-coding RNA(lncRNA) RP11-770J1.3 and transmembrane protein 25 (TMEM25) on paclitaxel resistance in human breast cancer MCF-7/PR cell line.

METHODSThe expression of lncRNA RP11-770J1.3 and TMEM25 in human breast cancer MCF-7(paclitaxel sensitive) and MCF-7/PR(paclitaxel resistant) cells were detected by quantitative RT-PCR. The synthetic interfering fragments of lncRNA RP11-770J1.3 and TMEM25 were transfected into MCF-7/PR cells. Sulforhodamine B assay was used to detect the sensitivity of MCF-7/PR cells to paclitaxel after interference of lncRNA RP11-770J1.3 and TMEM25. The expression of multidrug-resistance genes and proteins were detected by qRT-PCR and Western blot, respectively.

RESULTSlncRNA RP11-770J1.3 and TMEM25 were highly expressed in MCF-7/PR cells, and were significantly down-regulated after transfection of synthetic interfering fragments. Down-regulation of lncRNA RP11-770J1.3 and TMEM25 enhanced the sensitivity of MCF-7/PR cells to paclitaxel, and inhibited the expression of MRP, BCRP and MDR1/P-gp (all<0.05). Such effects were more significant when lncRNA RP11-770J1.3 and TMEM25 were both down-regulated (all<0.05).

CONCLUSIONSlncRNA RP11-770J1.3 and TMEM25 are highly expressed in MCF-7/PR cells, and the down-regulation of lncRNA RP11-770J1.3 and TMEM25 can enhance paclitaxel sensitivity in MCF-7/PR cells.

OBJECTIVETo investigate the effect of CXC chemokine receptor 4 (CXCR4) on cell cycle of breast cancer and its molecular mechanisms.

METHODSThe expression of CXCR4 and S phase kinase associated protein 2 (Skp2) was detected by real-time fluorescence quantitative PCR (fqRT-PCR) and Western blot in breast cancer cells. The expression of signal proteins and the downstream genes of Skp2 was detected by Western blot. The effect of CXCR4, PI3K/Akt pathway inhibitor LY294002 and ERK pathway inhibitor U0126 on cell cycle of breast cancer was detected by propidium iodide staining.

RESULTSSkp2 was significantly down-regulated in CXCR4-downregulated cells and up-regulated in CXCR4-upregulated cells. CXCR4 also regulated the expression of Skp2 and other downstream genes by signaling protein. The proportion of cells in G/Gphase increased and that in S phase declined in CXCR4-downregulated cell, and the effect was more significant when combined with the use of LY294002 or U0126.

CONCLUSIONSCXCR4 can affect cell cycle and inhibit the proliferation of breast cancer cells by regulating Skp2 gene expression through PI3K/Akt and ERK signaling pathway.

Objectives To access the bacteriology in patients with sepsis due to biliary tract infection to provide a basis for empirical selection of proper antibiotic treatment.Methods This is a single-center retrospective study on 214 patients with biliary tract infection admitted from August 2014 to July 2016 to the surgical intensive care units (ICU) of The First Affiliated Hospital of Sun Yat-sen University.To study the demographic information,sequential organ failure assessment (SOFA),usage of antibiotics before ICU and duration of ICU were analyzed.Bile,peritoneal drainage and blood samples were collected.Results 47 septic shock patients and 25 septic patients due to biliary tract infection were enrolled in the trial.The two groups (the shock group vs.the sepsis group) had a significant difference in the duration of ICU stay [(6.4 ± 4.6) d vs.(2.3 ± 1.8) d,P ＜ 0.05].48 strains of pathogens were isolated from the bile samples.The major pathogens were Escherichia coli (E.coli) (n =23,47.9％),Enterococcus faecalis (n =8,16.7％) and Enterococcus faecium (n =2,4.2％).80 strains of pathogens were isolated from the peritoneal drainage culture samples.E.coli,pseudomonas aeruginosa,and Klebsiella pneumoniae ranked the top 3 species,accounting for 26.3％,11.3％ and 7.5％,respectively.The sensitivity of E.coli isolated from bile to amikacin,imipenem and panipenem were all over 90.0％.Conclusions E.coli was the principal gram-negative bacterium in biliary infection induced sepsis.Early administration of carbapenemes may reduce the occurrence of septic shock in these patients.