1.Application of new generation high-throughput RNA sequencing in quality control of live attenuated yellow fever vaccine(chicken embryo cell) virus seed bank
Chinese Journal of Biologicals 2023;36(11):1335-1340
Objective To perform quality control in live attenuated yellow fever vaccine(chicken embryo cell)virus seed bank at the genomic level using the new generation Illumina/Solexa sequencing platform.Methods The live attenuated yellow fever vaccine strain YF17D-204 was inoculated into primary chicken embryo cells,and the chicken embryo cell adapted strains of live attenuated yellow fever vaccine were screened to establish YFV17D-CEC tertiary virus seed bank. The genome RNA of virus seeds was extracted,and the RNA library was prepared. The new generation Illumina/Solexa sequencing platform was used for high-throughput RNA sequencing. The whole genome nucleic acid sequence of yellow fever virus was systematically analyzed by using biological softwares such as FastQC,Trimmomatic,SPAdes,GapFiller,PrInSeS-G,Prokka,RepeatMasker,CRT,NCBI Blast~+,KAAS,HMMER3,TMHMM,SignalP,LipoP,ProtCamp and MegAlign.Results The whole genome of YFV17D-CEC tertiary virus seed bank contained 10 862 nucleotides,including an open reading frame(ORF)from 119 to 10 354(10 236 bp),encoding 3 412 amino acids. Sequence alignment analysis showed that the sequence of YF17D-CEC tertiary virus seed bank was 100% identical with YFV17D RKI(JN628279.1),YF/Vaccine/USA/Sanofi-Pasteur-17D-204/UF795AA/YFVax(JX503529.1)and YFV17D-204(KF769015.1),and no mutation occurred in the whole genome of the tertiary virus seed bank. Comparison of the sequences of different live attenuated yellow fever vaccine strains showed that yellow fever virus had multiple polymorphic sites.Conclusion YFV17DCEC has good genetic stability in primary chicken embryo cells. High-throughput RNA sequencing technology can quickly detect the whole genome information of YF17D-CEC virus seed bank,and the sequence analysis data can be used in the gene level quality control of yellow fever vaccine virus seed banks.
High-throughput RNA sequencing
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Live attenuated yellow fever vaccine
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Gene expression
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Virus seed bank
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Quality control
2.Importance of training on clinical thinking and clinical competence to interventional radiologists
Journal of Interventional Radiology 2006;0(08):-
Although the history of Interventional Radiology is no longer than 50 years,interventional techniques have been dramatically developed. Interventional radiologists have been responsible for much of the medical innovations and development of the minimally invasive procedures that are commonplace today to treat many complicated diseases as physicians. But the education backgrounds of interventional radiologist in China are different. Therefore,we should be aware that the job of an interventional radiologist is totally different from that of a diagnostic radiologist. It is very important to train interventional radiologists for improving their clinical thinking and clinical competence. Herein,we propose our suggestions on how to improve the clinical thinking and clinical competence of interventional radiologists. In this paper we also systemically introduce the accurate and proper treatment procedures which should be strictly followed in clinical work and,meanwhile,the perioperative patients care is emphasized.
3.Oinieal evaluation of interventional treatment for Budd-Chiari syndrome
Hongshan ZHONG ; Ke XU ; Liang XIAO
Chinese Journal of Radiology 2009;43(1):65-70
Objective To evaluate the interventional treatment of Budd-Chiari syndrome (BCS)with regard to different types of the disease.Methods One hundred and fifty-nine consecutive cases with BCS underwent interventional treatments with regard to different types of the diseases,including percutaneous angioplasty (PTA),transcatheter thrombolysis,endovascular stent implantation and modified transjugular intrahepatic portosystemic shunt(MTIPS).Among them,147 cases that underwent complete follow-up were enrolled in this study.Simple obstruction of HV,membranous obstruction of IVC,membranous obstruction of IVC combined with thrombosis in the distal lumen and segmental obstruction of IVC constituted 13.6/(20),66.0/(97),6.1/(9)and 14.3/(21/147),respectively.The technical success rate of each type Was determined.They were followed up for(67.3±9.0)months (16 h-104 months).Overall primary patency rote was evaluated.The late effect on liver function Was analyzed according to the Child-Push score.Results The primary patency rate of PTA was 65.6/ (86/131)and the secondary pateney rate was 96.9/(124/128).The primary patency rate of stent implantation was 78.9/(15/19)and the secondary patency rate Was 92.3/(24/26).One patient of type Ⅲa that received recanalization,catheter-directed thrombolysis and PTA in IVC died of hemoptysis 72 h after the procedtire.One patients of type I b who received MTIPS died of DIC 16 hrs after the procedure.And one patient of type Ⅳb who received MTIPS died of hver failure 13 months after the procedure.Twelve patients died in 7-79 months after the interventional procedure due to unrelated causes.At the end of follow-up,the liver function of the patients Was improved. Conclusions Optimal application of various vascular interventional techniques has a satisfactory primary and secondary patency rate and improves the long-term liver function.
4.Intensive care after transplantation
Youjun SUO ; Hongshan XU ; Li GONG
Chinese Journal of Tissue Engineering Research 2013;(44):7797-7802
BACKGROUND:It is an important and complex issue for the incidence of bacterial infection and complications after transplantation. The monitoring and care after transplantation can improve the success rate of transplantation. OBJECTIVE:To search the database of China National Knowledge Infrastructure, North American Clinical Trial Register and Thomson Reuters Web of Science database, and to perform literature metrological analysis and clinical trials registration project analysis on the published literatures of the monitoring and care after transplantation. METHODS:A total of 138 literatures were searched with the key words of“intensive care, transplantation”in the database of China National Knowledge Infrastructure on the intensive care after transplantation. 23 literatures were used for further analysis by reading titles and abstracts and 115 papers were excluded; the Thomson Reuters Web of Science database was searched with the key words of“intensive care, transplantation”for the literatures on the intensive care after transplantation published from 2008 to 2013;the North American Clinical Trial Register was searched with the key words of“intensive care, transplantation”for the related clinical trials, and a total of 50 registered projects were obtained, only 10 interventional studies. RESUTLS AND CONCLUSION:In recent years, the literatures on the intensive care after transplantation show a gradual increase trend. Compared with international research, fewer researches in this field emerge in China, literature quantity and quality need to be improved. There have been 1 693 papers published in the Thomson Reuters Web of Science database regarding the intensive care after transplantation. United States published the most literatures than other countries, total 532 papers, accounting for the largest proportion, 31.424%of total literatures. Transplantation Proceeding published the most literatures, total 144 papers, accounting for 8.506%of total literatures. There are 50 clinical trial registration projects related to the intensive care after transplantation in the North American Clinical Trial Register, 10 of them were interventional study, accounting for the majority, fol owed by observational study. Diagnostic study had no related registration projects. The surgery of transplantation was complex with trauma. The early monitoring and care post-transplantation are directly related to the success of surgery. Strengthening the monitoring and care of patients’ vital signs, rejection, bacterial infections and other aspects can reduce the complications and improve patients survival rate and quality of life.
6.Effect of angelica on myocardial fibrosis post myocardial infarction in rats
Yanfeng ZHAO ; Jiang XU ; Jianqing TANG ; Hong WANG ; Hongshan GUAN
Chinese Journal of Pathophysiology 2000;0(10):-
AIM: To investigate 1) the role of transforming growth factor-?_1(TGF-?_1) and macrophage infiltration during the development of myocardial fibrosis(MF) in rats after myocardial infarction(MI);and 2) mechanisms of MF post-MI and the inhibitory effect of angelica.METHODS: Sprague-Dawley(SD) rats were subjected to MI by ligating the left anterior descending coronary artery.The animals were randomly divided into three groups: sham,MI and MI+angelica.After 24 hours of ligation,rats received angelica(20 mL?kg~(-1)?d~(-1),ip) or saline.Left ventricular hemodynamics were measured and rats were killed at week 1,week 2 and week 4, respectively.Collagen content,macrophage infiltration and TGF-?_1 expression were examined in the non-infarcted area.RESULTS: ① In MI group,the numbers of macrophage and TGF-?_1 expression were significantly upregulated compared to sham at week 1 post-MI and remained elevated at week 4(P
7.Effective components against HIV-1 replicative enzymes isolated from plants.
Zonggen PENG ; Lijia XU ; Wencai YE ; Peigen XIAO ; Hongshan CHEN
Acta Pharmaceutica Sinica 2010;45(2):235-40
Plant active components characterized of many different structures and activities on multiple targets, have made them to be the important sources of inhibitors on HIV-1. For finding leading compounds with new structure against HIV-1, three key HIV-1 replicative enzymes (reverse transcriptase, protease and integrase) were used as screening models. The in vitro activities of 45 plant derived components isolated from Schisandraceae, Rutaceae and Ranunculaceae were reported. Within twelve triterpene components isolated, eight compounds were found to inhibit HIV-1 protease, in these eight active compounds, kadsuranic acid A (7) and nigranoic acid (8), inhibited both HIV-1 protease and integrase; Among fifteen lignans, meso-dihydroguaiaretic acid (15) and kadsurarin (16) were active on HIV-1 reverse transcriptase, and 4, 4-di(4-hydroxy-3-methoxyphenly)-2, 3-dimethylbutanol (13) active on HIV-1 integrase. All of the six alkaloids, seven flavones, and five others compounds were not active or only with low activities against HIV-1 replicative enzymes. Further studies of the triterpene components showing strong inhibitory activities on HIV-1 were warranted.
8.Significance of nonbronchial systemic artery embolization for massive hemoptysis
Shiping YU ; Ke XU ; Xitong ZHANG ; Hongshan ZHONG ; Xiuqin SU ; Jin ZHANG ; Tianyun MA
Chinese Journal of Radiology 2008;42(1):89-92
Objective To emphasize the importance of embolization of nonbronchial systemic arteries in treatment of acute and life-threatening massive hemoptysis.Methods In a series of 146 patients with hemoptysis who underwent bronchial artery embolization,we found 12 cases whose blood supply were from 17 nonbronchial systemic arteries and hemoptysis was more than 300 ml blood within 24 hours.Embolic materials included absorbable gelatin sponge(GS),kelp micro gelatin(KMG),polyvinyl alcohol(PVA) particles and metal coils. Results In the 12 cases with 17 nonbronchial systemic arteries (4 were intercostal,3 internal mammary,3 thyrocervical trunk,3 inferior phrenic,1 left gastric,2 originated from the inferior aortic arch,and 1 originated from anterior abdominal aortic wall).Five cases were embolized by GS alone,2 cases by KMG,3 cases by GS+PVA,and 2 cases by GS+PVA+metal coils.Eight cases were performed embolization once,3 cases were performed twice and 1 case was performed three times.No significant complications developed related to embolization,except that 1 patient had transient eyesight decrease after embolization of thyrocervical trunk and 2 patients had chest pain after embolization of intercostal artery which resovled without any treatment.Conclusions During bronchial artery embolization for hemoptysis patients,all supplying artery should be searched and found.Even after successful embolization of bronchial arterys for hemoptysis patients,nonbronchial systemic arterial supply should still be taken into account.
9.Mid-term follow-up of the percutaneous angioplasty of the occlusive aortoiliac artery diseases
Xitong ZHANG ; Hongshan ZHONG ; Liang XIAO ; Hongyi ZHANG ; Hoag LI ; Ke XU
Chinese Journal of Radiology 2009;43(4):415-417
Objective To evaluate the mid-term follow-up of the pereutaneous angioplasty of the occlusive acrtoiliac artery diseases.Methods The data of 30 patients who had distal abdominal aorta and bi-lateral iliac artery stenosis or occlusion and treated by percutaneous angioplasty were retrospectively reviewed.They are 24 males and 6 females,aged from 35 to 75 years (average 55 ± 10 years). The interventional procedures include wire-guided canalization,thrembolysis,balloon angioplasty and stents implantation.They were followed up by telephone,letter or visiting after discharged.Results Twenty-six patients with aarto-iliac are revascularized successfully.Three cases failed.Of them,one failed because only one side of iliac artery and aorta were successfully canalized while the eontralateral not,one failed to revascularize both the aorta and the iliac artery,the other one failed due to the ihac rupture and replaced by WallGraft implantation.Two cases had distal thrombosis during the procedure.One case was amputated due to the aggravated isebemia of the right lower extremity after the revascularization failed.Twenty-seven cases were followed uo after 1 to 112 months,averaged (41±9) months.One case showed that the iliac artery was uncanalized after 6 months.One case showed bilateral iliac artery occlusion during 25 months follow-up and re-occluded 3 months after balloon angioplasty and finally had stent implanation in bilateral iliac artery.The other patients had no aggravated symptom of recurrence.Conclusion The percutaneous angioplasty is a good choice for occlusive aorto-iliac artery disease,showing good recovery during mid-term follow-up.
10.Preparation and identification of monoclonal antibodies against human programmed cell death 10(PDCD10)
Yaoyao CHEN ; Yungang ZHAO ; Ting ZHANG ; Lanjun XU ; Xi MA ; Hongshan ZHAO
Journal of Peking University(Health Sciences) 2003;0(06):-
Objective: To obtain monoclonal antibodies against programmed cell death 10(PDCD10) for further study of the structure and function of PDCD10 protein.Methods: Balb/c mice were immunized with recombinant PDCD10,hybridoma cell lines secreting monoclonal antibodies against PDCD10 were screened by regular cell fusion and subcloning approach.The specificities of these monoclonal antibodies were determined by ELISA,Western blotting and Immunofluorescecence assay.Results: Three hybridoma cell lines(5G1,4F7 and 3H5) stable in secreting specific monoclonal antibodies were successfully obtained.Subclass of IgG belonged to IgG1(4F7 and 5G1)and IgG2b(3H5),respectively.The ascite titers of these monoclonal antibodies reached 1∶10~7.They could specifically bind to recombinant PDCD10 and endogenous and overexpressed PDCD10 proteins proved by ELISA and Western blotting.They failed to react with E.coli lysates and glutathione S-transferase(GST).In addition,these three monoclonal antibodies could recognize different epitopes of PDCD10 proteins assessed by immune fluorescence competitive binding assay.Both endogenous and overexpressed PDCD10 protein mainly located in the nucleus.Conclusion: Monoclonal antibodies against PDCD10 with high titers and specificity have been successfully prepared,which has laid the foundation for further study of PDCD10 protein.