2.Production and Identification of Broad Spectrum Monoclonal Antibody against a Group of Pyrethroid Insecticides
Mengtang WEN ; Yuan LIU ; Shuai YAN ; Xiao ZHANG ; Heng WANG ; Xianjin LIU
Chinese Journal of Analytical Chemistry 2014;(9):1245-1251
The objective of this study is to generate broad spectrum monoclonal antibody ( mAb ) against a group of pyrethroid insecticides and to identify its immunological characteristics. The generic hapten 3-phenoxy-benzoic acid ( PBA) was conjugated to carrier protein BSA by activated ester method. Balb/c mice were immunized with PBA-BSA. The titer of polyclonal antibody ( pAb ) was detected by indirect enzyme-linked immunosorbent assay ( ELISA) after five times immunization. The mouse with high titer and sensitivity was selected for cell fusing. The splenocytes of immunized mice were fused with Sp2/0 cells and the cultural supernatants of hybridoma cells were screened by indirect non-competitive ELISA based on the coating antigen PBA-ovoalbumin ( PBA-OVA ) . High-sensitivity and high-specificity mAb was prepared after subcloning using limiting dilution method. Purified mAb was obtained after purified by saturated ammonia sulfate precipitation and protein G affinity column. The immunological characteristics of mAb such as titer, antibody subtypes, affinity constant and the sensitivity to pyrethroid insecticides were characterized by indirect ELISA; The results of UV spectroscopy and SDS-PAGE showed that PBA-BSA artificial antigen was synthesized successfully. A hybridoma cell line (4H11) secreting anti-pyrethroid mAb was established. The titre of ascites was up to 1:6. 5×106, and the mAb was IgG1 subtype. The affinity constant of the mAb to PBA was about 2. 5×107 L/mol, with a IC50 value of 208. 83 μg/L and a detection limit of 21. 23 μg/L to PBA. Simultaneously, beta-cypermethrin, flucythrinate, cypermethrin and fenvalerate were sensitively recognized by the mAb with the IC50 of 1. 01, 2. 15, 3. 16 and 3. 67μg/L, respectively.
3.Prevalence of food intolerance and its related factors among health check-up receivers
Youfu CHENG ; Ping SHUAI ; Yuping LIU ; Hua YANG ; Fan YANG ; Rong ZHOU ; Li ZHANG ; Xian XIAO
Chinese Journal of Health Management 2012;06(5):311-314
Objective To study the prevalence of food intolerance and to explore its related factors among adult health check-up receivers.Methods A total of 863 adults who took physical examinations in our hospital from April to October 2011 were enrolled in this investigation.Height,body weight and blood pressure were measured,and serum IgG level was measured by enzyme-linked immunosorbent assay (ELISA).Results The total positive rate of food intolerance was 73%,and the leading intolerance items were crab (40.1% ),egg (29.8% ),cod fish ( 21.6% ),milk ( 20.0% ) and soybean ( 14.4% ).Females showed significantly higher prevalence of food intolerance than males.Various positive rate of milk or soybean intolerance was found in different age groups.No correlations of serum specific IgG with body mass index and systolic or diastolic blood pressure were observed.In logistic regression analysis,the odds ratio of food intolerance of women was 1.67 ( 95 % confidence interval 1.190 to 2.607 ).ConclusionsThe prevalence rate of food intolerance was high.The risk for food intolerance was significantly increased in women.Specific IgG antibody detection may help to early prevent and diagnose food intolerance-related diseases.
4.Pandanus tectorius derived caffeoylquinic acids inhibit lipid accumulation in HepG2 hepatoma cells through regulation of gene expression involved in lipid metabolism.
Chong-ming WU ; Hong LUAN ; Shuai WANG ; Xiao-po ZHANG ; Hai-tao LIU ; Peng GUO
Acta Pharmaceutica Sinica 2015;50(3):278-283
The fruit of Pandanus tectorius (PTF) has a long history of use as a folk medicine to treat hyperlipidemia in Hainan province, South China. Our previous studies have shown that the n-butanol extract of PTF is rich in caffeoylquinic acids and has an adequate therapeutic effect on dyslipidemic animals induced by high-fat diet. In this work, seven caffeoylquinic acids isolated from PTF were screened for the lipid-lowering activity in HepG2 hepatoma cells. Oil-Red O staining, microscopy and intracellular triglyceride (TG) and total cholesterol (TC) quantification showed that 3-O-caffeoylquinic acid (3-CQA), 3, 5-di-O-caffeoylquinic acid (3,5-CQA), and 3,4,5-tri-O-caffeoylquinic acid (3,4,5-CQA) significantly inhibited lipid accumulation induced by oleic acid and decreased intracellular levels of TC and TG in a dose-dependent manner. These three caffeoylquinic acids showed no significant cytotoxicity at concentrations of 1 -50 μmol x L(-1) as determined by MTT assay. Realtime quantitative PCR revealed that 3-CQA and 3, 5-CQA significantly increased the expression of lipid oxidation-related genes PPARα, CPT-1 and ACOX1 while 3-CQA, 3, 5-CQA and 3,4,5-CQA decreased the expression of lipogenic genes SREBP-1c, SREBP-2, HMGR, ACC, FAS. Overall, 3-CQA, 3, 5-CQA and 3, 4, 5-CQA may be the principal hypolipidemic components in PTF which can decrease intracellular lipid accumulation through up-regulating the expression of lipid oxidative genes and down-regulating the expression of lipogenic genes.
Carcinoma, Hepatocellular
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metabolism
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China
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Cholesterol
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metabolism
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Gene Expression Regulation
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Hep G2 Cells
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Humans
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Lipid Metabolism
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Liver Neoplasms
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metabolism
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Oleic Acid
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Pandanaceae
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chemistry
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Quinic Acid
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analogs & derivatives
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chemistry
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Sterol Regulatory Element Binding Protein 1
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Triglycerides
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metabolism
5.ZHU Lian--the founder of Chinese acupuncture-moxibustion scientific research.
Yang-Shuai SU ; Bing LIU ; Xiang-Hong JING ; Wei HE ; Xiao-Yu WANG ; Li-Jian ZHANG
Chinese Acupuncture & Moxibustion 2014;34(12):1221-1224
This article discussed ZHU Lian's contributions to acupuncture-moxibustion scientific research from three aspects: building the scientific thought of "new acupuncture-moxibustion", constructing the first domestic acupuncture-moxibustion institution and opening the door to modern acupuncture-moxibustion scientific research. ZHU Lian's visionary thought of "new acupuncture-moxibustion" has influenced the following researchers till now. She established the acupuncture-Moxibustion therapeutic institute affiliated to the Ministry of Health, set up the acupuncture-Moxibustion research platforms and teams and made research cooperation. She firstly carried out acupuncture-Moxibustion clinical and basic scientific research, which started the acupuncture-Moxi- bustion scientific research in China. ZHU Lian is the Pioneer of Chinese acupuncture-Moxibustion scientific research.
Acupuncture
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education
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Acupuncture Therapy
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history
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China
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History, 20th Century
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Humans
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Moxibustion
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history
6.3D-TV health assessment system by the multi-modal physiological signals.
Zhongqiang LI ; Lidong XING ; Zhiyu QIAN ; Xiao WANG ; Defei YU ; Baoyu LIU ; Shuai JIN
Chinese Journal of Medical Instrumentation 2014;38(2):84-87
In order to meet the requirements of the multi-physiological signal measurement of the 3D-TV health assessment, try to find the suitable biological acquisition chips and design the hardware system which can detect different physiological signals in real time. The systems mainly uses ARM11/S3C6410 microcontroller to control the EEG/EOG acquisition chip RHA2116 and the ECG acquisition chip ADS1298, and then the microcontroller transfer the data collected by the chips to the PC software by the USB port which can display and save the experimental data in real time, then use the Matlab software for further processing of the data, finally make a final health assessment. In the meantime, for the different varieties in the different brain regions of watching 3D-TV, developed the special brain electrode placement and the experimental data processing methods, then effectively disposed the multi-signal data in the multilevel.
Computer Simulation
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Equipment Design
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Microcomputers
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Physiological Phenomena
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Signal Processing, Computer-Assisted
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instrumentation
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Software
7.Expression of transcriptional factor Snail and adhesion factor E-cadherin in rectal cancer
Shuai XIAO ; Zhengjun QIU ; Chen HUANG ; Feng LI ; Longfei LIU ; Xianzhou LU
International Journal of Surgery 2010;37(8):514-519,封3
Objective To investigate the expression of E-cadherin and Snail proteins in rectal cancer and their significance. Methods The expression of Snail and E-cadherin proteins was detected using immunohistochemical SABC method in 101 cases of rectal cancer tissues. Results The positive rate of Snail in rectal cancer was 78.2% (79/101). The negative expression rate of E-cadherin in rectal cancer was 62.4% (63/101). The expression of Snail and E-cadherin were significantly related with the lymph node metastasis and Dukes' stage of rectal cancer (P < 0.05). Conclusion The overexpression of Snail and the decreased expression of E-cadherin might be important biological markers for malignant transformation, invasion and metastasis of rectal carcinoma.
8.Implantation of a triamcinolone acetonide drug delivery system into the suprachoroidal space for the prevention of traumatic anterior proliferative vitreoretinopathy
Shuai, QIN ; Zhong-qiu, JIN ; Xiao, CHEN ; Bo-chuan, WANG ; Hong, LIU
Chinese Journal of Experimental Ophthalmology 2012;30(5):428-432
BackgroundAnterior proliferative vitreoretinopathy (aPVR)is a tissue injury and repair progress,and treatment of aPVR is very important in clinic.Chitosan drug delivery system is becoming a hot spot for its large lading dose and long acting duration.ObjectiveThe present study was to investigate the curative effect of a triamcinolone acetonide (TA) drug delivery system after implantation into the suprachoroidal space to treat traumatic aPVR.MethodsaPVR models were created in the left eyes of 65 healthy pigment rabbits by performinga 5 mm penetrating incision 2.5 mm posterior to limbum at 10:30-11:30.The animals were randomly divided into 4groups.Blank chitosan was implanted into the suprachoroidal space as the blank control group.Chitosan with 1 mg TA was implanted in the TA + chitosa group.The TA solution ( containing 1 mg TA) was intravitreally injected in the TA injection group.Fifteen models were used as the traumatic control group.Another 15 left eyes of normal pigment rabbits were used as the normal control group.The thickness of the ciliary tissue was measured using a ultrasound biomicroscope(UBM) 3,5 and 8 weeks after operation.The animals were sacrificed by excessive anesthesia and eyeballswereobtainedforhistopathologicalandultrastructuralexaminations.ResultsHistopathological examination showed the edema of the ciliary tissue and inflammatory cells infiltration in the blank control group,TA injection group and model control group,but mild response was seen in the TA + chitosa group.Severe damage in the ciliary tissue and subcellular organelle was found in the blank and model control groups,but mild damage was detected in the TA + chitosa group under the transmission electron microscope.UBM examination revealed that obvious abnormalities were visible in the ciliary and iris tissue in the blank control group,TA injection group and traumatic control group,but a mild abnormality was seen in the TA + chitosa group.Significant differences in ciliary thickness were exhibited among the 5 groups 2,5 and 8 weeks after operation (F =212.938,515.323,447.919,P<0.01 ).Compared with the normal control group,ciliary thickness significantly increased in the blank control group and normal control group at various time points (all P<0.05 ),but that in the TA + chitosa group was significantly lower than the normal control group at various time points ( two weeks:0.484±0.075 vs.0.327 ±0.094 ; five weeks:0.422 ±0.089vs.0.327±0.094 ;eight weeks:0.418±0.085 vs.0.327±0.094) (all P>0.05). ConclusionsThe chitosan drug delivery system with TA suppresses the excessive proliferation of injured ocular tissue after implantation into the suprachoroidal space,which prevents the formation and development of aPVR.
9.Purification of HLA-DR molecules
Shuai-Zheng, JIA ; Hong-Yan, SUN ; Xiao-Da, LIU ; Zhi-Yan, DU ; Quan-Li, WANG
Bulletin of The Academy of Military Medical Sciences 2001;25(1):13-16
Objective:To purify HLA-DR molecules. Methods: Anti-HLA-DR antibody L243 was purified and coupled with CNBr activated Sepharose 4B gel. Immunoaffinity column was used to purify HLA-DR molecules. Results:Twenty micrograms of HLA-DR molecules were isolated from about 5 g Epstein-Barr virus-transformed human B lymphoblastoid cell line RAJI lysates by affinity chromatography. The purified HLA-DR molecules existed in α/β heterodimers form and could bind to conformation-dependent antibody L243. These HLA-DR molecules were separated into two strands,α and β,by boiling denaturation. These results are the basis for studying MHC Ⅱ binding peptide motif and CD4+ T cell epitopes of antigens in future.
10.Expression and clinical significance of aldehyde dehydrogenase 1 in nasopharyngeal squamous carcinoma
Fengjiao YANG ; Xiangwei WU ; Shuai XIAO ; Cuihong JIANG ; Lili HE ; Jingshi LIU
Cancer Research and Clinic 2013;25(10):675-677,680
Objective To investigate the significance of aldehyde dehydrogenase 1 (ALDH1)expression in appraising the prognosis of nasopharyngeal squamous carcinoma (NPC).Methods The expression of ALDH1 protein was detected by immunohistochemistry in human specimens obtained from 120 NPC patients without the history of radiotherapy or chemothetapy.Results A relatively high expression of ALDH1 were observed in 40.0 % (48/120) patients of NPC,the expression of ALDH1 in the edge of the cancer nests and stroma,particularly spindle cancer cells were strongly positive.The positive expression of ALDH1 was closely associated with patients' nasopharyngeal tumor size (P =0.011),lymphatic metastasis (P =0.005) and clinic stage (P =0.001),but not associated with their gender and age (both P > 0.05).The Kaplan-Merier and Cox regression analysis indicated that ALDH 1 closely correlated with clinical progrosis of the patients.It showed that ALDH1 was an independent risk factor which may affect the prognosis of NPC patients.Conclusion The expression of ALDH1 protein closely correlate with clinic outcome of NPC,suggesting that ALDH1 is a risk factor for clinic progrosis of NPC patients.