1.Cloning and Transcriptional Activity Analysis of Endogenous U6 Promoters in Artemisia annua
Yuting PU ; Bohan CHENG ; Mengyue WANG ; Jun ZOU ; Ranran GAO ; Lan WU ; Qinggang YIN ; Li XIANG ; Yuhua SHI
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(24):161-167
ObjectiveThe U6 promoter is an essential element for driving sgRNA expression in the clustered regularly interspaced short palindromic repeat sequences/CRISPR-associated protein 9(CRISPR/Cas9)gene editing system in dicotyledonous plants. Endogenous U6 promoters typically exhibit higher transcriptional activity, which can significantly improve gene editing efficiency. This study aims to identify endogenous U6 promoters in Artemisia annua to optimize its CRISPR/Cas9 gene editing system, which holds significant importance for its molecular breeding. MethodsOn the basis of the highly conserved U6 snRNA sequences in Arabidopsis thaliana, endogenous U6 promoters were screened in the A. annua genome. Expression vectors were constructed with candidate AaU6 promoter driving the firefly luciferase (LUC) reporter gene, and then transiently transformed into Nicotiana benthamiana. Transcriptional activities of the promoters were measured and compared by in vivo imaging and the Dual Luciferase Reporter assay. ResultsEight endogenous U6 promoters were successfully cloned from A. annua. Sequences alignment revealed that all these promoters contained the two conserved cis-acting elements, upstream sequence element (USE) and TATA-box, which affected their transcriptional activity. Dual-luciferase activity assays indicated that the transcriptional activities of AaU6-3, AaU6-1, and AaU6-5 were significantly higher than that of the Arabidopsis AtU6-26 promoter, with AaU6-3 exhibiting the highest activity. ConclusionThis study identified three endogenous AaU6 promoters with high transcriptional activity in A. annua, providing key functional elements for establishing an efficient gene editing system in A. annua. These findings will contribute to advancing precision molecular breeding and high-quality germplasm innovation in A. annua.
2.Molecular Mechanism and Therapeutic Exploration of CD36 in Breast Cancer
Shengqiao FU ; Qian JI ; Xinyu SUN ; Xi PU ; Yuting WU ; Haowei TANG ; Wanying SHENG ; Xu WANG
Cancer Research on Prevention and Treatment 2024;51(5):380-385
Breast cancer is the most diagnosed cancer in women worldwide and the leading cause of most cancer-related deaths,posing a serious threat to women′s health worldwide.At present,although the prognosis of some patients with breast cancer has improved,the emergence of drug resistance and the metastasis and recurrence of breast cancer are still the main reasons for poor prognosis.CD36 is a multiligand transmembrane glycoprotein expressed on various cell types.In recent years,studies have confirmed that CD36 can reshape the lipid metabolism of cancer cells;promote the differentiation of tumor-related macrophages into M2 type and recruitment into tumor tissues;regulate the function of Treg cells,CD8+T cells,DCs,and other immune cells,and thus promote tumor development.In addition,CD36 is also associated with breast cancer stem cells,metastasis-initiating cells,and breast drug resistant cells.Therefore,CD36 could be an important potential therapeutic target for breast cancer.
3.Analysis of clinical characteristics and prognosis of 4 264 patients with asymptomatic and mild novel coronavirus infections in Shanghai
Xiangru XU ; Ding SUN ; Min CAO ; Wen ZHANG ; Yuting PU ; Caiyu CHEN ; Yuting SUN ; Shuang ZHOU ; Bangjiang FANG
Chinese Critical Care Medicine 2022;34(5):449-453
Objective:To analyze the clinical characteristics and prognosis of patients infected with novel coronavirus Omicron variant in Shanghai, as to provide a reference for epidemic prevention, clinical diagnosis, and treatment.Methods:Altogether 4 264 novel coronavirus Omicron variant-infected patients with positive results of nucleic acid admitted to Shanghai New International Expo Center N3 Mobile Cabin Hospital from April 2 to May 7, 2022, were included. The demographic and baseline clinical characteristics, treatment strategy, prognosis, and different factors affecting the length of hospital stay were analyzed.Results:A total of 4 264 novel coronavirus variant Omicron-infected cases were collected, including 3 111 cases (73.0%) asymptomatic infections and 1 153 cases (27.0%) mild infections. The overall median age was 45 (33, 55) years old with a range from 2 years old to 81 years old. The male to female ratio was 1.37∶1. Altogether 3 305 cases (77.5%) had been vaccinated, of which 3 166 cases completed more than 2 doses. The upper respiratory tract symptoms such as cough and expectoration were the most common clinical manifestations of these infected patients. During the course of the disease, patients with asymptomatic infection were mainly treated with traditional Chinese medicine (TCM, 55.1%) and clinical observation (36.8%), and those with mild infection were mainly treated with TCM (42.2%) or integrated Chinese and Western medicine (30.4%). All patients were cured and discharged. The overall median length of hospital stay and the negative conversion time of nucleic acid were 9 (6, 10) days and 8 (5, 9) days, respectively. Compared with the asymptomatic infected patients, the hospitalization duration and the nucleic acid negative conversion time of the mildly infected patients were slightly longer [days: 10 (8, 11) vs. 9 (5, 10); 8 (6, 10) vs. 7 (4, 9), both P < 0.001]. Multiple linear regression analysis showed that the increasing age and mild infection were associated with longer hospitalization duration, and the treatment of TCM or integrated Chinese and Western medicine was associated with shortened length of hospital stay (all P < 0.05). Conclusions:The current novel coronavirus Omicron variant epidemic in Shanghai mainly caused asymptomatic and mild infections. The young and middle-aged population had a relatively high infection rate. The upper respiratory tract symptoms such as cough and expectoration were the most common clinical symptoms. Elderly and confirmed patients had prolonged hospitalization duration, while for patients receiving TCM treatment, the hospitalization duration was shortened.
4.Epidemiological and clinical characteristics of imported malaria in Dalian from 2013 to 2018
Changxin HAN ; Aijun SUN ; Yuting LI ; Fang SUI ; Shijuan QIN ; Chunwen PU
Chinese Journal of Endemiology 2019;38(10):835-839
Objective To understand the epidemiological and clinical characteristics of imported malaria cases admitted in Dalian and provide evidence for clinical diagnosis,treatment and control of the disease.Methods A retrospective analysis method was used to descriptively analyze the epidemiological data of 104 cases of imported malaria from 2013 to 2018 treated in Dalian Sixth People's Hospital.The clinical characteristics of 93 hospitalized patients (13 in the severe group and 80 in the non-severe group) were analyzed by t (t') test or Mann-Whitney U test.Results Among 104 cases of imported malaria,82 cases were falciparum malaria,5 cases were vivax malaria,4 cases were oval malaria,2 cases were quartan malaria,2 cases were mixed infections,and there were 9 cases without classification.The ratio of males to females was 16.33:1.00 (98:6).The age was (42.07 ± 11.07) years.There was no obvious seasonality in the onset time.We found 102 cases were come from Africa,and their main occupations were outbound workers or fishermen.After blood laboratory examination at admission between severe group and non-severe group,the differences of red blood cell (RBC),hematocrit (PCV),hemoglobin (Hb),serum creatinine (SCr),and blood urea nitrogen (BUN)were statistically significantly different (t =6.561,7.140,6.962;Z =-3.469,-3.739,P < 0.05).Conclusions In Dalian the falciparum malaria is the main infectious species in imported malaria cases,and Africa is the main area of infection.Outbound workers should be trained in malaria prevention and treatment in Africa.Early admission indicators (RBC,PCV,Hb,SCr,BUN) help clinicians to diagnosis and treat severe cases early.

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