1.In vitro induction of p210~(Bcr-Abl) protein-specific cytotoxic T cell responses using 562 cell total RNA transfected human dendritic cells
Li GAO ; Huahua FAN ; Huazhong LU
Chinese Journal of Blood Transfusion 1988;0(04):-
Objective To investigate whether human monocyte-derived dendritic cells (DCs) can express p210 Bcr-Abl protein and induce antigen-specific CTL responses in vitro after transfection with total RNA of K562 cells (K562-RNA).Methods Immature DCs were derived from human peripheral blood monocytes after 5 day incubation in the presence of GM-CSF and IL-4. The cells were then transfected with K562-RNA using electroporation or DOTAP lipofection. To verify the successful transfection of DCs with K562-RNA, Bcr-Abl fusion gene expression was determined by RT-PCR and Western blot. The immune phenotypes of the DCs were analyzed by flow cytometry. CTL cytotoxicity was assayed by propidium iodide (PI) stain and flow cytometry. The amount of DCs, CD1a expression and purity of DCs were measured by FACS.Results Bcr-Abl fusion gene appeared in the DCs after transfection with K562 cell total RNA. But 24 hours later, the Bcr-Abl mRNA from the K562-RNA transfected DCs disappeared, while the cells were expressing p210 Bcr-Abl protein. The transfected DCs could significantly promote T lymphocytes to kill the target K562 cells. We found that PBMC can be induced to DC in culture medium containing human plasma, suggesting a potential for clinical application.Conclusion Human dendritic cells transfected by K562 total RNA can induce effective p210 Bcr-Abl protein-specific immune responses, which might broaden the spectrum of possible DC-based clinical applications.
2.Clinical analys of esophageal cancer patients treated by combined composite Kushen injection and radiotherapy
Ping CAI ; Qingshen LIU ; Huahua CHEN ; Yixuan LI ; Xihui LI
Cancer Research and Clinic 2000;0(06):-
Objective To analyze the efficacy and debase the toxicity of radiotherapy combined with composite Kushen injection in treatment of esophageal carcinoma patients. Methods A total of 126 cases of esophageal cancer were treated by radiotherapy were analysed. All patients were treated with the continuative hyperfractionated radiotherapy. The total dose was 66 ~ 74 Gy/5 ~ 7 wk, 2.2 ~ 2.4 Gy/d of esophageal carcinoma patients. Two groups were given respectively delivered with the same radioactive dosages and methods.But the synthetic group was treated by radiotherapy with composite Kushen injection. composite Kushen injection was given at the dosage of 20 ml with 500 ml of saline infusion for 20 consecutive days. Results X-ray grades of esophageal cancer after radiotherapy. The grade I of X-ray manifestation after radiotherapy for patients in synthetic group was superior to that patient in the radiotherapy alone group. (60.4 % vs 41 %; ?2 =4.57, P
3.Inhibition of melanoma cell proliferation by targeting Wnt/β-catenin pathway through Sox4 RNA interference.
Huahua, CAI ; Anhong, NI ; Wen, LI ; Jiawen, LI
Journal of Huazhong University of Science and Technology (Medical Sciences) 2011;31(4):565-9
The effect of siRNA-mediated Sox4 gene silencing on Wnt/β-catenin signaling pathway of human malignant melanoma cell line A375 was investigated. Two types of dsRNA targeting Sox4 were constructed and transfected into A375 cells, and untreated cells and cells transfected with scramble RNA were used as blank control and negative control respectively. The expression levels of mRNA and protein of Sox4, Wnt3a, β-catenin and Wnt/β-catenin signaling target gene Survivin were detected after real-time PCR and Western blot respectively. MTT assay was used to measure cell proliferation after Sox4 knockdown. β-catenin/TCF transcription reporter assay was used for assessing Wnt/β-catenin signaling pathway activity. Our results showed that the two types of Sox4 siRNA were transfected into A375 cells successfully. As compared with untreated cells, Sox4 siRNAs had no significant influence on Wnt3a expression, and Sox4 siRNAs led to the decrease of β-catenin at protein level. Wnt/β-catenin signaling pathway activity was inhibited significantly. As a target of Wnt/β-catenin signaling, Survivin was decreased at both mRNA and protein levels, and cell proliferation was attenuated. Our study suggests that Sox4 may play an important role in Wnt/β-catenin signaling pathway in human malignant melanoma cells by regulating β-catenin protein level, indicating that Sox4 is involved in the progression of malignant melanoma through Wnt/β-catenin signaling pathway.
4.Analysis of Key Factors Affecting Maintenance Cost of Large-Scale Medical Equipment
Hanxue LI ; Wanhai QIAO ; Huahua ZHANG ; Huiru DOU
Chinese Medical Equipment Journal 2004;0(07):-
Objective To explore the key factors which affect maintenance cost of large-scale medical equipment. Methods The basic factors which affect maintenance cost of medical imagining equipment were summarized. The relationships of them are analyzed and then an interpretative structure mode of each factor was set. Additionally, the multiple relationship and controllability between each factor were also analyzed and the judging standard of key factors was set. Results The maintenance policy and mode were the key factors which affect maintenance cost of medical imagining equipment. Conclusion High quality maintenance for equipment condition and scientific optimization of the maintenance policy are necessary to fundamentally reduce the maintenance cost of large-scale medical equipment.
5.Research on the relationship between field cognition style and self-handicapping of medical students
Nina LI ; Huahua XU ; Feng JI ; Yu FENG ; Weigong XIE
Chinese Journal of Behavioral Medicine and Brain Science 2014;23(6):550-552
Objective To explore the relationship between field cognitive style and self-handicapping of medical students.Methods 124 medical students were measured by embedded figure test (EFT) and self-handicapping questionnaire (SHS).Results ①The scores of self-handicapping questionnaire were significantly different in whether they like their major or not(33.61±5.21,35.94±5.16,t=-0.218,P=0.031).Differences were also found in whether they could obtain the scholarships (33.20±6.19,35.12±4.12,t=-0.247,P=0.043),and in whether they were in a relationship(35.35±5.48,33.39±5.01,t=2.063,P=0.041).②Compared with field-independent cognitive style students,Field-dependent cognitive style students ' self-handicapping score was higher (35.69±3.91,32.63±6.08,t=3.303,P=0.001).③Field-dependent cognitive style,dislike their major and be in a relationship were the significant predictors of self-handicapping.Conclusion Correlation exists between field cognitive style and self-handicapping of the medical students.Students who belong to field-dependent cognitive style are more prone to self-handicapping than field-independent cognitive style students.
6.Development of Portable Automatic Tourniquet
Wei LI ; Zuyun ZHONG ; Mingfang ZHOU ; Qin SHU ; Huahua YIN
Chinese Medical Equipment Journal 1989;0(02):-
Objective To develop a portable automatic tourniquet.Methods Using microcomputer and pressure sensor,the pressure and hemostatic time of current pressurized tourniquet were controlled.Results Portable automatic tourniquet was composed of gasbag pressurized bandage,electric micro-pump,pressure sensor,preamplifier,impact damper,A/D converter,single-chip micro-computer controller,data memory,keyboard and its interface circuit,display and its interface circuit,photoelectronic control circuit,etc.Conclusion Portable automatic tourniquet is small,light,safe and convenient.Besides,rapid hemostasia and automatically controlled & stable pressure enable it to be used both in the war and at peace time.[Chinese Medical Equipment Journal,2008,29(2):21-23]
7.Prediction of outcomes in patients with acute basilar artery occlusion using 24 h NIHSS score after endovascular treatment
Yan LI ; Lei LIU ; Huahua LI ; Zongen GAO
International Journal of Cerebrovascular Diseases 2023;31(4):241-247
Objective:To investigate the predictive value of the National Institutes of Health Stroke Scale (NIHSS) score at 24 h after endovascular treatment on the outcomes in patients with acute basilar artery occlusion (ABAO).Methods:Consecutive patients with ABAO received endovascular treatment at the Department of Neurology, Shengli Oilfield Central Hospital from January 2019 to December 2020 were retrospectively included. According to the modified Rankin Scale scores at 90 days after onset, the patients were divided into a good outcome group (0-3) and a poor outcome group (4-6), as well as a survival group and a death group. The demographic and clinical data between the groups were compared respectively. Multivariate logistic regression analysis was use to identify independent influencing factors for clinical outcomes and mortality. The predictive value of postprocedural 24 h NIHSS score on the outcomes was evaluated using the receiver operating characteristic (ROC) curves. Results:A total of 35 patients with ABAO were included. Their age was 62 years (interquartile range, 56-66 years), and 28 patients were males (80%); 19 (54.3%) had a good outcome, 16 (45.7%) had a poor outcome, and 7 (20.0%) died. Univariate analysis showed that there were statistically significant differences in hypertension, low-density lipoprotein cholesterol, fasting blood glucose, collateral circulation grading, vascular recanalization, and postprocedural 24 h NIHSS scores between the good outcome group and the poor outcome group (all P<0.05). Multivariate logistic regression analysis showed that the postprocedural 24 h NIHSS score was independently correlated with the poor outcome (odds ratio 1.131, 95% confidence interval 1.017-1.258; P=0.023). Multivariate analysis did not find the independent influencing factors for death. ROC curve analysis showed that the area under the curve of the postprocedural 24 h NIHSS score for predicting poor outcome was 0.814 (95% confidence interval 0.668-0.960; P=0.011). The optimal cutoff value was 19 points, and the corresponding sensitivity and specificity were 85.7% and 71.4% respectively. Conclusions:In patients with ABAO receiving endovascular treatment, the postprocedural 24 h NIHSS score has good predictive value for poor outcomes at 90 d after procedure.
8.Construction of controlled expression system of class B G-protein coupled receptor PAC1.
Mei LI ; Rongjie YU ; Jiaping ZHONG ; Zekai CUI ; Yanxu YANG ; Huahua ZHANG
Chinese Journal of Biotechnology 2014;30(4):636-643
PAC1 is the neuropeptide pituitary adenylate cyclase activating polypeptide (PACAP) preferring receptor, which belongs to class B G protein-coupled receptors (GPCR) family. PAC1 mediates the most effects of PACAP as neurotransmitter, neuroregulator and neuroprotectant, while its high expression has close relationship with some physiological and pathological processes such as nerve-injury and tumor. To further understand the function of PAC1, a cell line that expressed inducible PAC1 was constructed to achieve Doxycycline (Dox) dependent expression of PAC1 in CHO (Chinese hamster ovary) cell using the improved Tet (tetracycline)-on Advanced System. First, the PAC1-EYFP fusion gene composed of PAC1 gene and gene encoding EYFP (enhanced yellow fluorescent protein) was sub-cloned to the tetracycline response element pTRE-Tight vector to construct the recombinant vector pEYFP-PAC1-EYFP by double enzyme digestion. Second, the tetracycline regulation components pTet-On advanced vector and the response element pTRE-PAC1-EYFP vector were both introduced into CHO cells successively and the positive clones were screened with G418 and hygromycin respectively. Third, the controlled expression of PAC1-EYFP in CHO was induced by tetracycline analogues Dox in different concentrations and the different levels of receptor PAC1-EYFP were detected. The results of fluorescence analysis and western blotting show that the cell strain with Dox dependent expression of PAC1-EYFP named PAC1-Tet-CHO was obtained. Moreover, in PAC1-Tet-CHO cells the expression of PAC1-EYFP was induced by Dox in a dose-dependent manner. The inducible expression of PAC1 still was stable after sub-culturing for more than 10 passages. It was also found by MTT assay that the higher expression level of PAC1 endowed the cells with higher proliferative viabilities. The construction of controlled expression system of PAC1 will lay a foundation for the further research on PAC1 profiles.
Animals
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Blotting, Western
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CHO Cells
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Cloning, Molecular
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Cricetinae
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Cricetulus
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Genetic Vectors
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Receptors, Pituitary Adenylate Cyclase-Activating Polypeptide, Type I
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biosynthesis
9.An empirical study on medical selection of flying cadets with hernia disease between PLAAF and USAF
Hao LI ; Huahua WEI ; Xueming WEI ; Haiyang ZHOU ; Zhongli MA ; Keshun ZHU ; Jian WANG ; Zhikang ZOU
Military Medical Sciences 2016;40(9):755-758
Objective To analyze the disease spectrum of abdominal external hernia during medical selection of flying cadets and compare the medical standards between US Air Force and Chinese PLA Air Force in order to improve the system of medical selection.Methods The data about abdominal external hernia among flying cadets during Chinese PLAAF medical selection between 2012 and 2015 were retrospectively analyzed and compared with the medical standards of the USAF flying cadets.Results Twenty-nine candidates were eliminated because of hernia,including 4 cases of hiatal hernia,3 cases of umbilical hernia and 22 cases of inguinal hernia.4.70% of the candidates were eliminated because of hernia disease.Empirical research showed difference in hernia standards between China and the USA.Conclusion Compared with Chinese medical selection standards,the USAF standards give more weight to whether the symptoms of disease impact air safety and efficiency.Our medical selection standards need to be improved by learning from foreign practices.
10.Correlation of onset time and influencing factors for ischemic stroke and myocardial infarction
Huahua LI ; Ke DENG ; Yan LI ; Xuemei LI ; Zongen GAO
International Journal of Cerebrovascular Diseases 2023;31(5):390-394
Cerebrovascular diseases and cardiovascular diseases are among the top causes of death worldwide. In recent years, the concept of "brain-heart co-treatment" has gradually gained popularity. Cerebrovascular and cardiovascular diseases have similar pathological foundations, pathogenesis, comorbidities, and treatment methods. However, there is currently limited research on whether the two have a chronological order of the first onset and which is the index event of the other. However, elucidating the above issues is of great significance for the secondary prevention of cardiocerebrovascular diseases. This article reviews the correlation of onset time and influencing factors for ischemic stroke and myocardial infarction.