1.Prokaryotic expression of human cTnI and preparation of anti-cTnI monoclonal antibody
Chinese Journal of Laboratory Medicine 2003;0(11):-
Objective To prepare monoclonal antibody (mAb) against recombinant human cardiac troponin I(cTnI).Methods The full-length gene encoding human cardiac troponin I (cTnI) was synthesized chemically and inserted into expression plasmid pBV220 to construct recombinant plasmid p pBV220/cTnI. The recombinant plasmid was transformed into E.coli DH5? which then expressed cTnI. The immunological activity of the expressed cTnI was analyzed by Western blot. Recombinant human cTnI protein was used as antigen to immunize BALB/c mice. Monoclonal anti-bodies against cTnI were prepared by normal hybridoma technology. The relative affinity of mAbs was determined by ELISA. Specificity of mAbs was analyzed by Western blot.Results Human cTnI gene was synthesized and confirmed by DNA sequencing. Positive recombinant clones were identified by restriction enzyme digestion analysis and DNA sequencing. Western blot analysis showed that the cTnI protein could be recognized by an anti- cTnI antibody. Two hybridmas producing antibodies against cTnI were obtained. IgG isotypes of two mAbs were IgG2a and IgG2b. Western blot showed that the antibodies were specific for cTnI. Neutralisation test showed that these mAbs could be evidently neutralized by cTnI.Conclusion The recombinant expression plasmid of cTnI was constructed successfully and expressed in E.coli. The method of EL ISA established to test serum cTnI is to clinically useful. The cTnI mAb which using cTnI as antigen prepared in this paper can be used for cTnI immunoassay in vitro.
2.Study on Rules for Acupoint Selection of Warming Acupuncture and Moxibustion in Treatment of Knee Osteoarthritis Based on Data Mining
Neng CHEN ; Xuemeng XU ; Wengang LIU ; Deming ZENG ; Guocai CHEN
Chinese Journal of Information on Traditional Chinese Medicine 2016;23(3):31-34
Objective To analyze the rules for acupoint selection of warming acupuncture and moxibustion in clinical treatment of knee osteoarthritis based on data mining technology. Methods CNKI, Wanfang Data, CBM, VIP, PubMed and Cochrane Library were searched for relevant literature about warming acupuncture and moxibustion in treatment of knee osteoarthritis. Database was built according to the search results. Association rules were used to analyze the rules for acupoint selection. Results Totally 189 papers were included, involving 189 prescriptions of acupuncture and moxibustion, 46 acupoints, 1270 times of acupoint selection. There were 1119 times of neighboring acupoint selection (88.1%), 151 times of distant acupoint selection (11.9%), 654 times of acupoint selection in yang meridian (51.5%) and 283 times of acupoint selection in yin meridian (22.3%). Conclusion Warming acupuncture and moxibustion in clinical treatment of knee osteoarthritis focuses on acupoints in Stomach Meridian of Foot-Yangming, Spleen Meridian of Foot-Taiyin; the neighboring acupoint selection is the main method, combined with evidence-based distant acupoint selection; acupoint selection from yang meridian was emphasized.
3.THE IMPACTS OF FOLIC ACID, RIBOFLAVIN AND MTHFR C677T POLYMORPHISM ON HUMAN LYMPHOCYTES GENOMIC STABILITY
Xu WANG ; Xiaoling XIA ; Xiayu WU ; Xiangxu KAN ; Neng CAO
Acta Nutrimenta Sinica 2004;0(05):-
Objective: To explore the comprehensive effects of folic acid (FA), riboflavin (RF) and MTHFR C677T polymorphism on genomic stability. Method: Cytokinesis-block micronucleus assay was used to detect the effects of different concentration combination of FA (20 and 200nmol/L, i.e. LF and HF) , RF (1 and 500 nmol/L, i.e. LR and HR) and MTHFR C677T polymorphism on genomic stability of 9 d cultured human lymphocytes. Results: The genetic damage was significantly higher in LFHR group regardless the genotype (P
4.Severe complications after microwave ablation in 7 403 cases with liver cancer
Yanming WANG ; Neng WANG ; Yun XU ; Qiang SHEN ; Guojun QIAN
Chinese Journal of Hepatobiliary Surgery 2016;22(10):655-660
Objective To identify the types,frequency and risk factors of major complications using microwave ablation (MWA) to treat liver malignancies in a single center.Methods Using the electronic medical record system,patients with liver cancer who were treated with MWA from January 2010 to November 2015 were retrospectively studied,and the risk factors of severe complications were analyzed.Results Of the 7 403 patients who were treated with MWA (12 558 ablations).96 cases experienced severe complications (1.3%) and 5 patients died (0.07%).The complications included liver abscess (n = 34,0.46%),pleural effusion (n = 20,0.27%),bile duct injury (n = 15,0.20%),hemorrhage (n = 6,0.08%),liver failure (n = 6,0.08%),extrahepa-tic organ injuries (n = 5,0.07%),cancerous cells implantation of abdominal walls (n = 2),cardiac arrest (n = 1),vascular thrombosis (n = 1),renal insufficiency (n = 1),and patients with other associated diseases which developed after the MWA therapy (n =2).Metastatic liver cancer and earlier treatment of MWA sessions were associated with a higher major complication rate (P < 0.05).Conclnsions MWA is a well-tolerated local treatment for liver cancer,with an acceptably low incidence of severe complications.Liver abscess was the most common postoperative severe complication.The surgeons' experience and tumor type were associated with the incidence of severe complications,however,not relavant with the frequency of preoperative ablations.Appropriate measures can be taken to effectively reduce the risk of serious complications.
5.A Clinical and Experimental Study on the Use of Decoction of Jie Du Hua Yu Tang in the Treatment of Psoriasis
Shengli WU ; Neng XU ; Jianming CHEN ; Yongmei LI ; Shaoyao MA
Chinese Journal of Dermatology 1995;0(03):-
Objectives To study the clinical effects and therapeutic mechanisms of the application of Chinese traditional medicine,the Decoction of Jie Du Hua Yu Tang,in the treatment of psoriasis.Meth-ods The clinical effects were observed in50patients with psoriasis vulgaris,to whom the Decoction of Jie Du Hua Yu Tang was given.The mice were used as experimental models whose tail scale epidermis was nat-urally lacking granular layer,similar to psoriatic epidermis.The mice were fed with Jie Du Hua Yu Tang and the effects on the formation of granular layer in their tail scale epidermis were noticed.Results The clinical cure rate was42%and the total response rate was90%with the Jie Du Hua Yu Tang.There was a signifi-cantly strong promoting effect on the formation of granular layer in mouse tail scale epidermis with the use of Jie Du Hua Yu Tang,in comparison with the control group.Conclusions The Decoction of Jie Du Hua Yu Tang is effective for the treatment of psoriasis vulgaris.The decoction could promote the formation of granu-lar layer in mouse tail scale epidermis.The therapeutic mechanism of the decoction may be related to the inhibition of proliferation of epidermal cells.
6.Efficacy analysis of percutaneous microwave ablation for patients with unresectable early stage hepatocel-lular carcinoma
Neng WANG ; Qiang SHEN ; Yun XU ; Guojun QIAN
The Journal of Practical Medicine 2016;32(13):2164-2167
Objective To investigate the efficiency and prognostic factors for the patients with hepatocel-lular carcinoma (HCC) at unresectable early stage when treated with percutaneous microwave ablation. Methods From January 2007 to January 2010 , 105 cases of at unresectable early stage were treated with percutaneous microwave ablation. Complete response rate, major complication rate, overall survival rates, disease-free survival rate were observed and prognostic risk factors were analyzed. Results The complete response rate was 95.7%. The major complication ratewas 3.8%. The 1-, 3-, and 5-year overall survival rates were 97.1%, 66.7%, 46.7%, and the corresponding disease-free survival rates were 82.9%, 48.6%, 31.4%, respectively. Age > 65 year and tumor size > 3 cm were independent risks for disease-free survival. Age > 65 year and recurrence were independent risks for overall survival. Conclusions Percutaneous microwave ablation is a safe and efficient ther-apy for HCC at unresectable early stage.
7.Clinical analysis on second poisoning in medical staff rescuing patients with acute phosgene poisoning.
Neng-Luan XU ; Yu-Sheng CHEN ; Zhang-Shu LIN
Chinese Journal of Industrial Hygiene and Occupational Diseases 2006;24(12):738-738
Adult
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Female
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Humans
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Male
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Medical Staff
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Middle Aged
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Occupational Exposure
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Phosgene
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poisoning
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Poisoning
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etiology
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Young Adult
8.THE IMPACTS OF VARIATION OF FOLATE METABOLIC COMPONENT AND MTHFR C677T POLYMORPHISMS ON CHROMOSOME SEGREGATION
Juan NI ; Lin LU ; Neng CAO ; Tao ZHOU ; Weijiang XU ; Xu WANG
Acta Nutrimenta Sinica 2004;0(06):-
Objective To study the impacts of variation of folate metabolic component including folic acid (FA), riboflavin (RF) and MTHFR C677T polymorphism on chromosome 8 and 17 segregation. Method RPMI1640 medium was modified with different combinations of low (LF) and high (HF) concentrations of folic acid (20, 200 nmol/L) and low (LR) and high (HR) concentrations of riboflavin (1 and 500 nmol/L). The lymphocytes with different MTFHR C677T genotype were cultured in the mentioned media for 9d. Cytokinesis was blocked by cytochalasin B and the slide was prepared as usual. The frequencies of aneuploidy of chromosomes 8 and 17 were determined by dual-color fluorescence in situ hybridization (FISH). Results The frequencies of both chromosome aneuploidy were significantly higher in low FA (LFLR+LFHR) than that in higher FA (HFLR+HFHR) (P
9.Recent development of natural and reconstituted lipoprotein based nano drug delivery vehicles.
Ying XU ; Xue-Feng JIN ; Qi-Neng PING ; Hong-Fei LIU ; Mei CHEN ; Xi-Ming XU
Acta Pharmaceutica Sinica 2014;49(1):23-29
Lipoproteins are biological lipids carriers. The natural and reconstituted lipoprotein based drug delivery systems have been extensively developed in recent years. This article reviews the development of natural and reconstituted low-density lipoprotein and high-density lipoprotein based vehicles in the antitumor area.
Animals
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Antineoplastic Agents
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administration & dosage
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chemistry
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Apolipoproteins B
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administration & dosage
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chemistry
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Drug Carriers
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administration & dosage
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chemistry
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Humans
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Lipoproteins
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administration & dosage
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chemistry
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Lipoproteins, HDL
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administration & dosage
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chemistry
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Lipoproteins, LDL
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administration & dosage
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chemistry
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Nanoparticles
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Neoplasms
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drug therapy
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Peptides
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administration & dosage
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chemistry
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Pharmaceutical Vehicles
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chemistry
10.Differentiation of mouse embryonic stem cells into insulin-secreting cells induced by a 5-step model system
Mu-Chao WU ; Hua CHENG ; Ming-Tong XU ; Li-Hong CHEN ; Feng LI ; Sheng-Neng XUE ;
Chinese Journal of Endocrinology and Metabolism 1986;0(03):-
Objective To induce mouse embryonic stem(ES)cells to differentiate into insulin-secreting cells by means of a 5-step model system.Methods E14.1 mouse ES cells were cultured in the presence of leukemia inhibitory factor(LIF)for 2 days(step 1),then the cells were cultured in hanging drops to form embryonic bodies(EBs)and the resulting EBs were cultured in suspension for 6 days in the presence of basic fibroblast growth factor bFGF(step 2).Subsequently the EBs were cultured in the medium containing glucagon- like peptide 1(GLP-1),hepatocyte growth factor(HGF),nerve growth factor(NGF)and nicotinamide for 10 days(step 3).After that,the EBs were dissociated into single cells,and the cells were cultured in monolayer in the presence of GLP-1,betacellulin,activin A,bFGF and nicotinamide for 10 days(step 4).Finally,the cells were cultured in low-glucose medium containing nicotinamide for 4 days(step 5).Insulin and some other islet- related genes expressions were investigated using RT-PCR and insulin expression was also investigated by DTZ- staining and immunohistochemistry.The percentage of insulin-secreting cells was evaluated by flowcytometry and insulin concentrations were measured by RIA.Results mRNA expression of insulin became visible at step 3 and more evident at step 5.Additionally,at step 5,mRNAs of glucagon,somatostatin,pancreatic polypeptide(PP), pancreatic duodenal homeobox 1(PDX-1),beta-cell E box transactivator 2(Beta2)and neurogenin 3(Ngn3) were detected.DTZ-staining positive cells and insulin immunohistochemical staining positive cells were observed. The percentage of insulin-positive cells was(24.0?2.5)%(n=6).In the presence of 5.6 mmol/L and 25 mmol/L glucose,insulin concentrations were(0.05?0.01)?g/L and(0.13?0.02)?g/L respectively(n= 6).Conclusion E14.1 mouse ES cells can be induced to differentiate into insulin-secreting cells by the 5-step model system.Insulin-secreting cells can release insulin into culture medium when treated with glucose,and insulin concentrations increase with rising concentration of glucose.