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.

Objective:To analyze the detection strategy and basic detection situation of markers of infectious diseases transmitted by transfusion in blood testing laboratories of blood stations in China.Methods:Based on the data of practice comparison working party of Blood Stations in Mainland of China from 2017 to 2021, the data on the testing strategies and the basic detection information of the markers for the transmission of infectious diseases through transfusion in the member laboratories of the practice comparison working party of Blood Stations in Mainland of China from 2017 to 2021 were collected, and the situation of the selection for testing markers, testing strategy and the testing method and other relevant aspects were sorted out and analyzed by charts.Results:The selection of the testing markers was consistent, but HTLV testing item was added in some member laboratories. The detection strategy of using two ELISA reagents and one nucleic acid testing (NAT) reagent simultaneously was adopted in 47 member blood stations; 3) NAT method was dominated by mini pool-NAT in member laboratories. The number of members adopting mini-pools of 8 (MP8)-NAT decreased from 17 in 2017 to 14 in 2021, while the number of members adopting mini-pools of 6 (MP6)-NAT increased from 13 in 2017 to 22 in 2021; Roche NAT system accounted for the largest proportion.Conclusions:In order to ensure blood safety and avoid missing detection, the blood stations still adopt the detection strategy of using two ELISA reagents and one nucleic acid testing (NAT) reagent simultaneously; Meanwhile, in order to increase the NAT positive rate, the proportion of mini pool-NAT mainly decreased year by year despite its dominating role, while the proportion of individual donation-NAT increased year by year; NAT method is transiting from mini-pools of 8 (MP8) to mini-pools of 6 (MP6); The proportion of imported NAT system used in NAT laboratory is relatively large.

Objective To study the effect of macrophage migration inhibitory factor(MIF) antibody on the rat colonic aberrant crypt foci(ACF) induced by 1,2-dimethylhydrazine(DMH),carcinoma number and expression of MIF in rat colonic carcinogenesis.Methods Rat colonic carcinogenesis model was induced by DMH.In this model,the inhibitory effect of MIF antibody on the number of ACF and carcinoma was observed.ELISA and immunohistochemical staining were adopted to investigate the effect of MIF antibody in early cancerative intestinal mucosa and MIF expression after cancer formation.Results The number of ACF and carcinoma was significantly inhibited by MIF antibody intervention(P＜ 0.01).The expression of MIF in the colonic carcinoma model was significantly higher than that in the pre-carcinoma ACF model(P＜0.01).Applying MIF antibody could significantly inhibit the expression of MIF in both rat colonic ACF and colonic carcinoma model.Conclusion MIF antibody can significantly inhibit the rat colonic mucosal carcinogenesis,which may be related with inhibiting number of ACF and expression of MIF.MIF antibody may be expected to become a new target spot of precaution and treatment of colonic carcinoma.

Objective To investigate the effect of simvastatin on prothrombotic state in OSAS patients with hypertension without hyperlipemia. Methods Eighty OSAS patients with hypertension without hyperlipemia were enrolled from Jan-uary 2011 to June 2014,and randomly divided into simvastatin group(40 cases)and conventional group(40 cases)(oral taken simvastatin or placeb once every night in addition of routine antihypertensive drugs).Forty healthy subjects were also recruited as control.All cases were followed up for eight weeks. Serum holesterol,nitric oxide(NO),emdothelin-1(ET-1),von Willebrand-factor(vWF) and tissue-type plasminogen activator(t-PA) levels were determined in each case before and after eight weeks orsuvastatin medication. Results Before treatment NO,t-PA levels in simvastatin group and conventional group were lower than the healthy group,ET-1, vWF levels were higher than those of control group,the differences were statistically significant(P<0.05). After treatment NO,t-PA levels in simvastatin group and conventional group increased compared with before treatment,but simvastatin group increased more significantly,ET-1,vWF levels in simvastatin group and conventional group were lower than before treatment,but simvastatin group they were more significantly lower, with statistically significant difference (P<0.05). Conclusion In patients of OSAS pa-tients with hypertension without hyper-lipemia,simvastatin can increase plasma NO,t-PA concentration and decrease ET-1,vWF concentration and thus to improve the prothrombotic status.

Objective To analyze the clinical diagnosis and treatment strategies of liver-localized posttransplantation lymphoproliferative disease (LL-PTLD).Methods Six cases of LL-PTLD from more than 3000 cases of liver transplant recipients from July 2003 to July 2011 were retrospectively analyzed.Other six cases of LL-PTLD were retrieved through Pubmed and Wanfang.The diagnosis and treatment of 12 cases of LL-PTLD were summarized and analyzed.Results All patients with LL-PTLD were diagnosed pathologically.The incidence of LL-PTLD was 0.2％ (6/3000).Among 12 patients,immunosuppressant and anti-EB virus treatment was reduced or withdrawn in the vast majority of patients,and treatment response was satisfactory.Systemic chemotherapy was given in 6 cases,and three of them died.Local radiation therapy was given in 4 cases,the tumor was significantly controled,and patients survived.Secondary liver transplantation was performed on 3 cases: 1 case died of recurrent lymphoma,and one case received partial hepatectomy and no lymphoma recurred.Conclusion For cases with obstructive symptoms of fever and chills associated with jaundice without reasonable explanation,LL-PTLD is suspected and diagnosed by liver biopsy.Basic treatments such as adjustment of immunosuppressive agents and anti-viral therapy are recommended as early as possible.Local radiation therapy is a treatment method of LL-PTLD,which can obtain a satisfactory therapeutic effect.

Objective To know the blood lead level of children in Handan city and the influence factors. Methods In Oct. 2004,211 children aged 2-6 years,living in Handan city for more than one year and without eliminating lead medicine treatment in recent three months,were chosen from a kindergarten. The blood lead was determined by WFC atomic absorption spectrometer. The influence factors were investigated by questionnaire.Results The blood lead level was(89.2?13.6) ?g/L,the prevalence of lead poisoning was 11.85%,no significant difference was seen between boys and girls. As for the blood lead level,the children living in the industrial areas or near traffic road was higher,the children living in the newly decorated house was higher,the children often eat preserved eggs was higher,the children whose parents engaged in the work of lead exposure was higher,the children whose parents smoke heavily at home everyday was higher,the children whose parents had lower education was higher. Conclusion More attention should be paid to prevent and control child lead poisoning in Handan city.