1.Enhancement of antitumor immunity of dendritic cells pulsed with hyperthermia lymphoma cells
Limei AI ; Jing PAN ; Yuanyuan SUN
Chinese Journal of Immunology 2010;26(3):232-235
Objective:To investigate the immuno-effects triggered by dendritic cells (DCs) pulsed with Heat-treated lymphoma cells.Methods:Mononuclear cells from healthy human peripheral blood(PBMC) were isolated and DCs were generated in vitro with normal methods.Lymphoma cells were heated (42℃,2 h) and then loaded onto DCs.Immunotypes of DCs and lymphocytes were detected with flow-cytometric analysis.IFN-γ releasing was detected by ELISPOT,lymphocyte cytotoxicity was analyzed by MTT method after mixing with DC reaction.Results:After pulsing DCs with hyperthermia lymphoma cells,CD80~+,CD86~+ and HLA-DR existed higher than those of the control group,IFN-γ releasing and cytotoxicities were also strengthened,but there was no difference between them,cytotoxitic lymphocytes (CTLs) were produced more as CD8~+,CD25~+ and CD56~+ cells by flow-cytometric analysis.Conclusion:Anti-lymphoma immuno-effects could be strengthened by loading DC with hyperthermia lymphoma cells.
2.Nosocomial Infection among Patients with Hematological Malignancies: A Retrospective Analysis
Limei AI ; Xin ZHANG ; Man YAN
Chinese Journal of Nosocomiology 2004;0(10):-
OBJECTIVE To prevent and control nosocomial infection(NI) among patients with hematological malignancies,and make clinical investigation.METHODS The random medical records of patients suffering from hematological malignancies in our hospital were analyzed retrospectively.RESULTS NI rate was 51.11%,in which the main infection site first was respiratory tract,next oral cavity and gastrointestinal tract.Gram-negative bacteria and fungi were the most common pathogens.CONCLUSIONS Patients with hematological malignancies are susceptible to NI,and they should be treated properly so as to prevent from infection effectively.
3.Study on immunological effect of dendritic cell transfected by recombinant plasmid ofΔNp73αon breast cancer
Yang WANG ; Xiaoqing MIAO ; Shuhua LI ; Limei AI ; Pei ZHANG
Chinese Journal of Immunology 2016;(2):169-173
Objective:To research the imumunological effect of dendritic cell transfected by recombinant plasmid of ΔNp73αon breast cancer.Methods:Cultivate dendritic cells by cytokines( GM-CSF,IL-4,TNF-α) from human umbilical cord blood in vitro, CD1a,CD83 were detected by flow cytometry.Recombinant plasmid pcDNA-HA/ΔNp73αwas used to transfected DCs,the transfection was detected by Western blot.Transfected DCs cocultivate with T cell induce specific cytotoxic lymphocytes( CTL).Proliferation ability of T cell was detected by MTT methods.IFN-γin the culture supernatant of the transfected DCs were detected by ELISA.The killing effect on breast cancer MDA-MB-231 was detected by LDH method.Results:The expression of CD1a(56%),CD83(74%)in mature DC was higher than immature DC(CD1a 19%,CD83 13%)(P<0.01).ΔNp73αtransfected group had a special band.The killing effect of specific CTL induced byΔNp73α-DC and T cells cocultivation on breast cancer cell MDA-MB-231 was statistic stronger than DC group(P<0.05),the proliferation of T cell and secretion level of IFN-γcompared with DC and pcDNA group were statistically significant(P<0.01).Conclusion:The CTL induced byΔNp73αtransfected DC vaccine has significant effect on killing breast cancer.
4.Correlation of Genomic Characteristic with Disease Progression in Polycythemia Vera
Yingxu ZHAO ; Jie BAI ; Lei ZHANG ; Mengyao SHENG ; Hui SHI ; Wen XING ; Fengchun YANG ; Limei AI ; Yuan ZHOU
Tianjin Medical Journal 2014;(6):517-521
Objective To screen mutations in genes including ASXL1, TET2, IDH1, IDH2, SETBP1, MPL515, JAK2 exon 12 and JAK2V617 in 135 polycythemia vera (PV) patients. To assess progreasson and genomics characteristics post polycythemic myelofibrosis. Methods DNA sequencing of ASXL1(Exon12),TET2 (Exons 3-11),IDH1(Exon4),IDH2(Ex-on4),SEPBP1(Exon4),JAK2 exon 12 and MPL515 (Exon 10) genes were carried out using Sanger method. JAK2V617 muta-tion was detected by allele-specific PCR in patients with PV. In the mean time, the mutation load of JAK2V617F allele (V617F%) was evaluated by real-time PCR using Tagman MGB probe. Then, the significant of gene mutations and clinical outcomes of post-PV Myelofibrosis(PPMF)was analyzed. To study risk factors of PPMF, logistic regression were employed. Results ASXL1, TET2, IDH1, IDH2 were mutated in 7.69%(8/104), 5.26%(1/19) , 0.08%(1/120) and 0.08%(1/121) of all PV patient respectively. JAK2 was mutated in 82.22%(111/135) of PV patients with mutation rate of exon12 of 2.96%(4/135) and there were no mutation of MPL515 and SETBP1 in PV patients. ASXL1 mutation was found in 31.82%(7/22) PPMF patients. Spearman analysis showed that ASXL1 is correlated with JAK2V617F (V617F%) (rs=0.298,P=0.002). The hemo-globin was lower in patients with ASXL1 mutation than patient without mutation (wild type). Leukocyte count, V617F%>50%rate, thrombosis and PPMF were higher in patients with ASXL1 mutation than that of ASXL1 wild type(P<0.05). ASXL1 mu-tation, V617F%>50% rate and splenomegaly were all risk factors of PPMF. Conclusion ASXL1 mutation is the risk-fac-tor of PPMF and may promote V617F%by some mechanism.
5.Estimation of the number of workers with occupational noise-induced hearing loss from manufacturing industry in China, 2020
Limei AI ; Xinxin LI ; Anqi LIU ; Dan WANG ; Jingxin WANG ; Weijiang HU ; Xia WAN
China Occupational Medicine 2023;50(5):512-517
{L-End}Objective To estimate the number of workers with occupational noise-induced hearing loss (ONHL) in the manufacturing industry of China in 2020. {L-End}Methods Multiple data sources were integrated to collect information on workers in the manufacturing industry from the Occupational Diseases and Hazards Monitoring Information System under China Disease Prevention and Control Information System. The total number of workers was split based on age from the 2018 Fourth National Economic Census and the 2020 National Survey of Occupational Disease Hazards. Additionally, data on the prevalence of hearing loss in the general population was adjusted based on the age composition of Jilin Province, and noise prevalence was standardized based on the age composition of the employed population in the Seventh National Population Censu. Attribution fractions (AF) was estimated. The prevalence ratio (PR) was calculated by the prevalence of hearing loss in the occupational noise-exposed workers and general population. The proportion and attributable cases (AC) of ONHL cases in all hearing loss cases were estimated for occupational noise-exposed workers. The number of ONHL was estimated based on AF and the total number of workers with hearing loss. {L-End}Results In 2020, a total of 30 059 525 workers were exposed to occupational noise in the manufacturing industry in China, with noise-exposed prevalence and standardized noise-exposed prevalence of 28.94% and 28.52%, respectively. The prevalence of hearing loss among occupational noise-exposed workers increased with age, and a total of 8 054 789 workers suffered from hearing loss. Most of the cases were at the age between 45-<55 years old. The total PR and 95% uncertainty interval (UI) was 2.83 (2.58-3.06) and the total AF and 95%UI was 64.63% (61.22%-67.30%), and both peaks were in the age of 30-<45 years. The AC and 95%UI was 5 205 980 (4 930 620-5 420 345) persons, and most of the cases were at the age between 40-<55 years. {L-End}Conclusion Occupational noise poses a serious threat to the hearing health of workers in the manufacturing industry of China, especially among middle-aged and young adults.
6. Clinical analysis of seven cases of H1N1 influenza-associated encephalopathy in children
Xufang LI ; Bin AI ; Jiawei YE ; Dengmin HE ; Limei TAN ; Minxia CHEN ; Huamei YANG ; Fansen ZENG ; Fengxia YANG ; Hongsheng LIU ; Yi XU
Chinese Journal of Pediatrics 2019;57(7):538-542
Objective:
To investigate the clinical manifestations, diagnosis, and treatment of H1N1 influenza A-associated encephalopathy (IAE) in children.
Methods:
The clinical manifestations, laboratory tests, cranial magnetic resonance imaging (MRI), electroencephalography (EEG) examinations and treatments of seven children with H1N1 IAE hospitalized in Guangzhou Women and Children′s Medical Center from December 2018 to January 2019 were retrospectively analyzed.
Results:
Five of the seven children with H1N1 IAE were female. The age at admission was 4 years and 5 months (range 7 months-9 years). Neurological symptoms occurred simultaneously or early (0-3 days) after the flu-like symptom appeared. The main clinical manifestations of neurological symptoms were seizures (repeated seizures in five cases and status convulsion in two cases, including one case of unexpected fever and repeated seizures in a nine-year old girl) accompanied with altered consciousness (drowsiness in five cases and coma in two cases). Cranial MRI in three cases displayed multifocal lesions, mainly in the bilateral thalamus, brainstem and cerebellar hemisphere. MRI also showed reversible splenial lesion in the corpus callusumin in three cases. EEG tracings were characterized by diffuse slow wave activity in four cases, and status epilepticus was monitored in one case. All the 7 cases were treated with oral oseltamivir. Three cases were treated with pulsed methylprednisolone and intravenous immunoglobulin. One case was treated with intravenous immunoglobulin alone and all the patients received oral oseltamivir. All the patients survived, with three patients had minor neurological sequelae at discharge.
Conclusions
The main clinical manifestations of H1N1 IAE are seizures and altered consciousness. Cranial MRI combined with EEG is helpful for early diagnosis. Intravenous immunoglobulin and (or) methylprednisolone should be considered for severe cases.
7.URA3 affects artemisinic acid production by an engineered Saccharomyces cerevisiae in pilot-scale fermentation.
Weiwei GUO ; Limei AI ; Dong HU ; Yajun CHEN ; Mengxin GENG ; Linghui ZHENG ; Liping BAI
Chinese Journal of Biotechnology 2022;38(2):737-748
CRISPR/Cas9 has been widely used in engineering Saccharomyces cerevisiae for gene insertion, replacement and deletion due to its simplicity and high efficiency. The selectable markers of CRISPR/Cas9 systems are particularly useful for genome editing and Cas9-plasmids removing in yeast. In our previous research, GAL80 gene has been deleted by the plasmid pML104-mediated CRISPR/Cas9 system in an engineered yeast, in order to eliminate the requirement of galactose supplementation for induction. The maximum artemisinic acid production by engineered S. cerevisiae 1211-2 (740 mg/L) was comparable to that of the parental strain 1211 without galactose induction. Unfortunately, S. cerevisiae 1211-2 was inefficient in the utilization of the carbon source ethanol in the subsequent 50 L pilot fermentation experiment. The artemisinic acid yield in the engineered S. cerevisiae 1211-2 was only 20%-25% compared with that of S. cerevisiae 1211. The mutation of the selection marker URA3 was supposed to affect the growth and artemisinic acid production. A ura3 mutant was successfully restored by a recombinant plasmid pML104-KanMx4-u along with a 90 bp donor DNA, resulting in S. cerevisiae 1211-3. This mutant could grow normally in a fed-batch fermentor with mixed glucose and ethanol feeding, and the final artemisinic acid yield (> 20 g/L) was comparable to that of the parental strain S. cerevisiae 1211. In this study, an engineered yeast strain producing artemisinic acid without galactose induction was obtained. More importantly, it was the first report showing that the auxotrophic marker URA3 significantly affected artemisinic acid production in a pilot-scale fermentation with ethanol feeding, which provides a reference for the production of other natural products in yeast chassis.
Artemisinins
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Fermentation
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Saccharomyces cerevisiae/metabolism*
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Saccharomyces cerevisiae Proteins/metabolism*