1. Metabolomics investigation on antiobesity effects of Corydalis bungeana on high-fat high-sugar diet-induced obese rats
Minghai FU ; LiSha A. ; Sungbo CHO ; Minghai FU ; Terigele BAO ; Hongzhen YU ; HuiFang LI ; Genna BA ; Sungbo CHO
Chinese Herbal Medicines 2022;14(3):414-421
Objective: Corydalis bungeana (CB) is a well-used medicinal herb in Mongolian folk medicine and has been traditionally applied as an antiobesity agent. However, the evidence-based pharmacological effects of CB and its specific metabolic alterations in the obese model are not entirely understood. This study aimed to utilize untargeted metabolomic techniques to identify biomarkers and gain mechanistic insight into the serum metabolite alterations associated with weight loss and lipid metabolism in obese rats. Methods: A high-fat high-sugar (HFHS) diet was used to induce obese models in rats. CB extract was orally gavaged at 0.18, 0.9 and 1.8 g/kg doses for six weeks, and feed intake, body weight, fat pad weight, and blood indexes were measured. Blood serum metabolites were evaluated by gas chromatography/quadrupole time-of-flight tandem mass spectrometry (GC-TOF/MS). Results: The results showed that compared with the obese group, the administration of CB extract caused significant decreases in body weight (P < 0.05), feed intake, Lee's index, and perirenal, mesenteric, epididymal fat weight. CB extract also reduced blood triglyceride and total cholesterol levels (P < 0.05) of obese rats. Metabolomic findings showed that nine differential metabolites, including pyruvic acid, D-glucuronic acid, malic acid, dimethylglycine, oxoglutaric acid, pantothenic acid, sorbitol acid, fumaric acid and glucose 6-phosphate were identified under CB treatment and altered metabolic pathways such as TCA cycle, pantothenate and CoA biosynthesis, and glycolysis/gluconeogenesis. Conclusion: This study demonstrated weight loss and lipid lowering effects of CB on HFHS diet-induced obese rats and identified nine metabolites as potential biomarkers for evaluating the favorable therapeutic mechanism of CB via regulation of lipid and glucose metabolism.
2.Development of a sandwich ELISA for detecting 3AB non-structural protein of foot-and-mouth disease virus.
Yuanfang FU ; Wei HE ; Pu SUN ; Lin YANG ; Huifang BAO ; Yimei CAO ; Xingwen BAI ; Pinghua LI ; Dong LI ; Yingli CHEN ; Lei LIU ; Zengjun LU ; Zaixin LIU
Chinese Journal of Biotechnology 2020;36(11):2357-2366
Antigenic purity is important for quality control of the foot-and-mouth (FMD) whole virus inactivated vaccine. The recommended method for evaluation the antigenic purity of FMD vaccine is to check the serum conversion to non-structural protein (NSP) 3AB antibody after 2 to 3 times inoculation of animals with inactivated vaccine. In this study, we developed a quantitative ELISA to detect the amount of residual 3AB in vaccine antigen, to provide a reference to evaluate the antigenic purity of FMD vaccine. Monoclonal antibody (Mab) of NSP 3A and HRP-conjugated Mab of NSP 3B were used to establish a sandwich ELISA to quantify the NSP 3AB in vaccine antigen of FMD. Purified NSP 3AB expressed in Escherichia coli was serially diluted and detected to draw the standard curve. The detectable limit was determined to be the lowest concentration of standard where the ratio of its OD value to OD blank well was not less than 2.0. Results: The OD value was linearly corelated with the concentration of 3AB protein within the range between 4.7 and 600 ng/mL. The correlation coefficient R² is greater than 0.99, and the lowest detectable limit is 4.7 ng/mL. The amount of 3AB protein in non-purified inactivated virus antigen was detected between 9.3 and 200 ng/mL depending on the 12 different virus strains, whereas the amount of 3AB in purified virus antigen was below the lowest detectable limit. The amount of 3AB in 9 batches of commercial FMD vaccine antigens was between 9.0 and 74 ng/mL, whereas it was below the detectable limit in other 24 batches of commercial vaccine antigens. Conclusion: the sandwich ELISA established in this study is specific and sensitive to detect the content of 3AB protein in vaccine antigen of FMD, which will be a useful method for evaluation of the antigenic purity and quality control of FMD inactivated vaccine.
Animals
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Antibodies, Viral
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Enzyme-Linked Immunosorbent Assay
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Foot-and-Mouth Disease/prevention & control*
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Foot-and-Mouth Disease Virus
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Viral Nonstructural Proteins/genetics*
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Viral Vaccines
3.Expression of long non-coding RNA 1010001N08Rik in bronchopulmonary dysplasia and its bioinformatics analysis
Tianping BAO ; Zhaofang TIAN ; Sai ZHAO ; Lijuan YANG ; Huaiping CHENG ; Yuan ZHANG ; Xiugui WANG ; Huifang WANG
Chinese Journal of Neonatology 2017;32(5):384-388
Objective To explore the expression feature of long non-coding RNA (lncRNA) 1010001N08Rik in hyperoxia-induced bronchopulmonary dysplasia (BPD) and predict the mechanism that 1010001N08Rik might be involved in the occurrence and development of BPD by a series of bioinformatics analysis.Method The sequence,genomic position and structure characteristics of 1010001N08Rik were acquired from UCSC genome browser,and its target gene was predicted by Ensemble database.We successfully established the animal model of BPD by making newborn C57BL/6J mice exposed to 95% concentrations of ambient oxygen for seven days.The expression of 1010001N08Rik and Gata 6 were detected using real-time quantitative polymerase chain reaction (PCR).Student's t test was used to compare their expression levels during the BPD process.Result The relative expression of 1010001N08Rik in BPD process at d1,d3,d5,d7 was 1.21 ± 0.33,2.02 ± 0.41,2.95 ± 0.45,4.20-± 0.48 respectively,and there were significant difference between adjacent time points (P < 0.05).The relative expression of Gata 6 mRNA was 0.92 ±0.30,1.10 ± 0.31,0.86 ± 0.24,0.45-± 0.08 respectively,and there was significant difference between d5 and d7 (P <0.05).1010001N08Rik had highly conserved property among different species.The chromosomal regions of 1010001N08Rik existed transcriptional factors binding locations and epigenetic regulation clues,and its possible candidate target gene was Gata 6.Conclusion The expression of 1010001N08Rik increased during the formation process of BPD.Bioinformatics analysis and preliminary experiment results suggested that 1010001N08Rik might participate in the process of BPD by down-regulating Gata 6 expression.
4.Effect of different noninvasive ventilation mode on the cardiac structure and function of patients with OSAHS
Yuexiang SHUI ; Haiyan TONG ; Shunjin ZHAO ; Xuxin BAO ; Huifang YANG ; Lixian TENG ; Ting YANG ; Maoxian ZHU ; Yang WU ; Xiaohong WU
Chinese Journal of Primary Medicine and Pharmacy 2016;23(13):1970-1974
Objective To compare the effect of continuous positive airway pressure (CPAP)and bilevel positive airway pressure(BiPAP)on the cardiac structure and function of patients with obstructive sleep apnea and hypopnea syndrome(OSAHS).Methods 100 patients with OSAHS were selected as the research subjects.The patients were randomly divided into two groups by digital table method,CPAP group had 50 cases,adopted the CPAP ventilation mode,BiPAP group had 50 cases,adopted the BiPAP ventilation mode.The heart rate,blood pressure, blood brain natriuretic peptide(BNP),nitric oxide(NO),endothelin -1 protease,matrix metalloprotein 9(MMP -9), C -reactive protein (CRP) and polysomnography (PSG) related indicators,cardiac structure and function and endothelial function changes before and after treatment were observed in the two groups,compared the clinical effect of the two modes.Results Before treatment,in both groups,there were no statistical differences in blood pressure,heart rate,BNP,NO,endothelin -1,MMP -9,CRP,PSG related indicators,cardiac structure and function,endothelial function.After six months of noninvasive ventilator treatment,results of the heart rate,systolic pressure,diastolic blood pressure,blood oxygen saturation,oxygen partial pressure,the BNP,endothelin -1,NO,MMP -9,CRP,body mass index,the Epworth sleepiness scale(ESS),Apnea hypoventilation index(AHI),oxygen and lowest at night,aortic di-ameter(AO),left ventricular inner diameter (LA),right ventricular(RV)inside diameter,left ventricular ejection fraction(LVEF),pulmonary artery systolic pressure(PASP)of the CPAP group were (79.83 ±11.47)times/min, (114.06 ±11.45)mmHg,(72.44 ±7.38)mmHg,(97.6 ±1.45)%,(93.17 ±1.86)mmHg,(110.78 ±38.32)ng/L, (17.58 ±2.07)ng/L,(8.55 ±0.55)μmol/L,(372.73 ±189.00)μg/L,(3.34 ±2.29)mg/L,(23.87 ±1.59), (0.98 ±0.70),(0.65 ±0.30),(94.04 ±1.62)%,(31.52 ±2.17 )mm,(31.19 ±1.09 )mm,(20.86 ± 1.69)mm,(61.13 ±5.02)%,(20.74 ±5.49)mmHg.which of the BiPAP group were (80.96 ±8.56)times/min, (114.58 ±9.34)mmHg,(71.67 ±8.57)mmHg,(96.96 ±1.43)%,(94.52 ±1.66)mmHg,(87.63 ±28.33)ng/L, (17.76 ±2.20)ng/L,(8.54 ±0.52)μmol/L,(359.63 ±268.95)μg/L,(4.96 ±2.00)mg/L,(24.15 ±1.65), (0.85 ±0.75 ),(0.58 ±0.19 ),(94.50 ±1.18)%,(31.73 ±1.57 )mm,(31.97 ±1.12)mm,(21.58 ± 2.43)mm,(62.24 ±5.79)%,(21.45 ±3.76)mmHg.In the oxygen partial pressure,the BNP,MMP -9,CRP,ESS score,AHI,LA,LVEF and other indicators,BiPAP mode were better than CPAP mode(t =2.13,4.32,2.13,4.32, 1.39,4.93,2.58,4.36,all P <0.05 ).Conclusion BiPAP mode and CPAP mode can improve cardiovascular function in patients with OSAHS,improve the symptoms of low ventilation,reduce obesity,but in terms of reducing cardiac load,improve blood vessel function,BiPAP mode is better than CPAP.
5.The molecular mechanism of interaction of trivalent dimethylarsinous acid (DMA(III)) binding to rat hemoglobin.
Min ZHANG ; Wenwen WANG ; Huifang JIN ; Lingling BAO ; Hua NARANMANDURA ; Yingjie QIN ; Chunhui LI
Acta Pharmaceutica Sinica 2014;49(5):666-71
In our previous work, we found that trivalent dimethylarsinous acid (DMA(III)) have high affinity binding to cysteine residue 13 of rat hemoglobin. However, it is still unknown why arsenic intermediate metabolite DMA(III) has high binding affinity for Cysl3 but not for other cysteine residues 93, 140, 111 and 125. In order to better understand the molecular mechanism of DMA(III) with rat hemoglobin, we have done current study. So, SD rats were divided into control and arsenic-treated groups randomly. Arsenic species in lysate of red blood cells were analyzed by HPLC-ICP-MS, and then determined by a hybrid quadrupole TOF MS. In addition, trivalent DMA(III) binds to different cysteine residues in rat hemoglobin alpha and beta chains were also simulated by Molecular Docking. Only Cys13 in alpha chain is able to bind to DMA(III) from the experiment results. Cys13 of alpha chain in rat hemoglobin is a specific binding site for DMA(III), and we found that amino acids compose pockets structure and surround Cys13 (but not other cysteine residues), make DMA(III) much easy to bind cysteine 13. Taken together, the DMA(III) specific binding to Cys13 is related to spatial structure of Cys13.
6.Enhanced immune response of a novel T-cell immunogen in vaccine for foot-and-mouth disease.
Qing ZHAO ; Pu SUN ; Zaixin LIU ; Pinghua LI ; Huifang BAO ; Yimei CAO ; Xingwen BAI ; Yuanfang FU ; Zengjun LU ; Dong LI
Chinese Journal of Biotechnology 2011;27(9):1281-1291
We investigated the enhanced immune response of a recombinant T cell immunogen as an effective cellular immune adjuvant. The T cell immunogen named TI contained several T cell epitopes from the VP1, VP4, 3A and 3D proteins of foot-and-mouth disease virus (FMDV) and two pan-T helper (T(H)) cell sites to broaden the immunogenicity of the protein. Meanwhile, another fusion protein named OA-VP1 was expressed in bacteria, which contained two VP1 proteins of O and Asia1 type FMDV. Mice were vaccinated with commercially inactivated vaccine or OA-VP1 protein with or without the TI immunogen. The results show that mice inoculated with inactivated vaccine or OA-VP1 protein supplemented with TI immunogen produced significantly higher level of neutralizing antibodies (P < 0.01 or P < 0.05) than the mice only inoculated with inactivated vaccine or OA-VP1 protein by microneutralization assay. An obvious increase in T cell number by flow cytometric analysis and significantly higher concentration of IFN-gamma secreted in culture media of spleen lymphocytes were observed in groups supplemented with TI immunogen (P < 0.01). TI immunogen was an effective stimulator for humoral and cellular immunity and could help improve the immunogenicity of inactivated vaccine or protein subunit vaccine.
Adjuvants, Immunologic
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pharmacology
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Animals
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Capsid Proteins
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genetics
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immunology
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Epitopes, T-Lymphocyte
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genetics
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immunology
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Foot-and-Mouth Disease
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immunology
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prevention & control
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virology
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Foot-and-Mouth Disease Virus
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immunology
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Immunization
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Mice
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Viral Vaccines
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genetics
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immunology
;
pharmacology
7.Application of SELDI-TOF-Ms technology in research of molecular markers of lung cancer
Huifang SHA ; Jianding YE ; Qiangling SUN ; Xiaohua YANG ; Guoliang BAO ; Jiuxian FENG ; Leluo GONG
Journal of Shanghai Jiaotong University(Medical Science) 2009;29(10):1178-1181,1195
Objective To explore the changes of proteomic spectra from plasma of patients with lung cancer or benign lung diseases and health controls in order to establish a primary diagnosis model of lung cancer. Methods The proteomic spectra from plasma of 108 patients with lung cancer, 40 patients with benign lung diseases and 22 healthy individuals were analysed by surface-enhanced laser desorption/ionization time of flight mass spectrometry ( SELDI-TOF-MS). The best decision tree model was established by cluster analysis and principal component analysis. Then the model was blindly validated by the protein of 21 patients with lung benign diseases and 47 patients with stage I lung cancer. Results Twenty-three significantly differentially expressed protein peaks were successfully detected (P <0.001). Blinded validation suggested that the accuracy for diagnosing lung cancer was 72. 06%, the sensitivity and specificity were 72. 34% and 71.43%, respectively, and the positive predictive value and negative predictive value were 85. 0% and 78. 95%, respectively. Conclusion SELDI-TOF-MS protein chip technology provides a new tool for the early diagnosis of lung cancer.
8.Amplification and Characterization of Bull Semen Infected Naturally with Foot-and-mouth Disease Virus Type Asial by RT-PCR
Junjun SHAO ; Huiyun CHANG ; Tong LIN ; Guozheng CONG ; Junzheng DU ; Jianhong GUO ; Huifang BAO ; Youjun SHANG ; Yamin YANG ; Xiangtao LIU ; Zaixin LIU ; Jixing LIU
Virologica Sinica 2008;23(5):378-382
To investigate the security of semen biologically, 15 bull semen samples were collected (of which 5 exhibited clinical signs of Foot-and-mouth disease) and identified by RT-PCR and virus isolation. The results indicated that the semen of the infected bulls were contaminated by Foot-and-mouth disease virus (FMDV), but FMDV was not detected in semen samples from those bulls not showing clinical signs of Foot-and-mouth disease (FMD). This is the first report of the presence of FMDV in bull semen due to natural infection in China. The analysis of the partial sequence of the VP1 gene showed that the virus strain isolated from semen has 97.9% identity with the virus isolated from vesicular liquid of infected bulls showing typical signs of FMD and belonged to the same gene sub-group.
9.Problem and Intervening for Ageing People with Intellectual Disabilities(review)
Huifang ZHANG ; Bing WANG ; Bao TIAN
Chinese Journal of Rehabilitation Theory and Practice 2008;14(5):419-421
Nowadays,the incidence of intellectual disabilities(ID) increased in the elder,and they would meet the following problems:life transitions,future life planning and understanding emotions,etc.This paper discusses the measures to be made for ageing person with ID from three aspects:assessing and managing illness,caregiving stress and the role of psychologists.
10.High Cell Density Culture of Lactobacillus plantarum Lp-2
Guangmin ZHANG ; Wei WANG ; Huifang BAO ; Xiaoling ZHANG ; Faqiang ZHAN ; Ruifeng LIN
China Biotechnology 2006;0(06):-
High cell density culture of Lactobacillus plantaru is a significant step of the preparation of lactic acid bacteria starter.Firstly,this experiment is designed to analyze the effects of different dissolved oxygen and pH on the batch fermentation of Lactobacillus plantarum.And then,on the base of the batch fermentation,fed-batch fermentation was studied,companied by chemic neutralization in the experiment to achieve the aim of high cell density cultivation.Finally,in the comparison of two kinds of feeding mode for fed-batch operation in 10L fermentor,it was demonstrated that two kinds of feeding mode made cell concentration increase differently,in which pH feedback feeding can control sucrose concentration within a certain lower range,therefore obtaining the maximum cell concentration.With the pH feedback feeding,the dry cell weight was up to 13.56g/L,which increased 90.05% over that of batch culture.


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