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abstract Cell metabolite analysis is of great interest to analytical chemists and physiologists, with some meta-bolites having been identified as important indicators of major diseases such as cancer. A high-throughput and sensitive method for drug metabolite analysis will largely promote the drug discovery industry. The basic barrier of metabolite analysis comes from the interference of complex components in cell biological system and low abundance of target substances. As a powerful tool in biosample analysis, microfluidic chip enhances the sensitivity and throughput by integrating multiple functional units into one chip. In this review, we discussed three critical steps of establishing functional microfluidic platform for cellular metabolism study. Cell in vitro culture model, on chip sample pretreatment, and microchip combined detectors were described in details and demonstrated by works in five years. And a brief summary was given to discuss the advantages as well as challenges of applying microchip method in cell metabolite and biosample analysis.
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Creatinine, uric acid, hypoxanthine and xanthine are important diagnostic biomarkers in human urine for gouty arthritis or renal disease diacrisis. A simple method for simultaneous determination of these biomarkers in urine based on reversed-phase high-performance liquid chromatography (RP-HPLC) with ultraviolet (UV) detector was proposed. After pretreatment by dilution, centrifugation and filtration, the biomarkers in urine samples were separated by ODS-BP column by elution with methanol/50 mM NaH2PO4 buffer solution at pH 5.26 (5:95). Good linearity between peak areas and concentrations of standards was obtained for the biomarkers with correlation coefficients in the range of 0.9957-0.9993. The proposed analytical method has satisfactory repeatability (the recovery of data in a range of creatinine, uric acid, hypoxanthine and xanthine was 93.49-97.90%, 95.38-96.45%, 112.46-115.78%and 90.82-97.13%with standard deviation of o5%, respectively) and the limits of detection (LODs, S/N Z 3) for creatinine, uric acid, hypoxanthine, and xanthine were 0.010, 0.025, 0.050 and 0.025 mg/L, respectively. The established method was proved to be simple, accurate, sensitive and reliable for the quantitation of gouty arthritis' biomarkers in human urine samples. The ratio of creatinine to uric acid was found to be a possible factor for assessment of gouty arthritis.
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A new modification method for glass slides was developed and applied to make ThinPrep Pap smears,in order to increase the adhesion ability of cervical exfoliative cells.3-glycidyloxypropyl trimethoxysilane (GOPS) was coated on the glass slides firstly on the slides,then poly-L-lysine (PLL)was covalently modified onto the above epoxy-terminated slides to form GOPS-PLL double decorated slides.The modified slides were characterized using X-ray photoelectron spectroscopy (XPS) and atomic force microscopy (AFM).The cell adhesion ability effect was tested and compared with traditional PLL coated slides by fixing the cervical exfoliative cells on the double adorned slides.The control test was conducted by the bare glass slides unmodified.The cell morphology of cervical exfoliative cells adhered on different slides was observed under the microscope after Papanicolaou staining.The number of cervical exfoliative cells on the unmodified slides,PLL coated slides and GOPS-PLL coated slides was 1030±300,3283±226 and 4119±280 (n=12),respectively.The data among the three different modification methods showed significant differences (one-way analysis of variance,ANOVA test,P＜ 0.05).The cell capturing effect of the GOPS-PLL slide was the best among the three different modified slides.In addition,the GOPS-PLL slide could enhance the uniformity of the adhered cells and be widely applied to the ThinPrep system for cervical carcinoma screening to increase the accuracy rate of diagnosis.
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A chemiluminescence enzyme immunoassay based on magnetic microparicles (MmPsCLEIA) was developed to evaluate serum α-fetoprotein (AFP) in parallel with tramional colorimetric enzyme-linked immunsorbrnt assay (ELISA).A sestematic comparison between the MmPs-CLEIA and colorimetric ELISA concluded that the MPs-CLEIA exhibited fewer of immunoreagents,less total assay time,and better linearity,recovery,precision,senitivity and validity.AFP was detected in forty human serum samples by the proposed MPs-CLEIA and ELISA,and the results werecompared with commercial electrochemilunminescence immunoassay (ECLIA) kit.The correlation coefficient between MPs-CLEIA and ELISA was obtained with R2=0.6703; however,the correlation between MPs-CLEIA and ECLIA (R2=0.9582) was obviously better than that between colorimetric ELISA and ECLIA (R2=0.6866).
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Glypican-3 （GPC3） is reported as a great promising tumor marker for hepatocellular carcinoma （HCC） diagnosis. Highly sensitive and accurate analysis of serum GPC3 （sGPC3）, in combination with or instead of traditional HCC marker alpha-fetoprotein （AFP）, is essential for early diagnosis of I-ICC. Biomaterial-functionalized magnetic particles have been utilized as solid supports with good biological compatibility for sensitive immunoassay. Here, the magnetic nanoparticles （MnPs） and magnetic microparticles （MmPs） with carboxyl groups were further modified with streptavidin, and applied for the development of chemiluminescence enzyme immunoassay （CLEIA）. After comparing between MnPs- and MmPs-based CLEIA, MnPs-based CLEIA was proved to be a better method with less assay time, greater sensitivity, better linearity and longer chemiluminescence platform. MnPs-based CLEIA was applied for detection of sGPC3 in normal liver, hepatocirrhosis, secondary liver cancer and HCC serum samples. The results indicated that sGPC3 was effective in diagnosis of HCC with high performance.
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Monolithic silica spin column extraction (MonoSpin-SPE) was developed as a simple,sensitive,and eco-friendly pretreatment method which combined with ultra-fast liquid chromatography-mass spectrometry (UFLC-MS) to determine the levels of six phthalate esters,dimethyl-(DMP),diethyl-(DEP),dipropyl-[DPrP],butyl-benzyl-(BBP),dicyclohexyl(DcHP),and di-n-octyl-(DOP) phthalate in physiological saline samples.Under optimized experimental conditions,the method was linear in the following ranges:0.2 - 50 μg/L for DMP,DEP,DPrP,DcHP and DOP; 5- 100,μg/L for BBP.The correlation coefficients (R2 ) were in the range of 0.9951- 0.9995 for all the analytes and the limits of detection (LODs) and limits of quantification (LOQs) were in the ranges of 0.02- 0.9 μg/L and 0.08- 2.7 μg/L,respectively.The pretreatment process showed good reproducibility with inter-day and intra-day relative standard deviations (RSDs) below 8.5％ and 11.2％,respectively.This method was used to determine the levels of six phthalate esters in physiological saline samples and the recoveries ranged from 71.2％ to 107.3％.DMP and DEP were found in actual physical saline samples (brand A and brand B).
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Glypican-3 (GPC3) is reported as a great promising tumor marker for hepatocellular carcinoma (HCC) diagnosis.Highly sensitive and accurate analysis of serum GPC3 (sGPC3),in combination with or instead of traditional HCC marker alpha-fetoprotein (AFP),is essential for early diagnosis of HCC.Biomaterial-functionalized magnetic particles have been utilized as solid supports with good biological compatibility for sensitive immunoassay.Here,the magnetic nanoparticles (MnPs) and magnetic microparticles (MmPs) with carboxyl groups were further modified with streptavidin,and applied for the development of chemiluminescence enzyme immunoassay (CLEIA).After comparing between MnPs- and MmPs-based CLEIA,MnPs-based CLEIA was proved to be a better method with less assay time,greater sensitivity,better linearity and longer chemiluminescence platform.MnPs-based CLEIA was applied for detection of sGPC3 in normal liver,hepatoeirrhosis,secondary liver cancer and HCC serum samples.The results indicated that sGPC3 was effective in diagnosis of HCC with high performance.
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In order to evaluate the moUuseicidal effect of 50% wettable power of niclosamide ethanolamine salt(WPN)combined with urea against Oncomelania snails in the field,4 g/m~2 WPN,4 g/m~2 WPN+20 g/m2 urea and 4 g/m~2 WPN+30 g/m~2 urea were used for mollusciciding with the spraying method.The results showed after 7 days,the mortality rates of snail were 74.43% for 4 g/m~2 WPN,90.32% for 4 g/m~2 WPN+20 g/m~2 urea and 94.83% for 4 g/m~2 WPN+30 g/m~2 urea,respectively.It is indicated that WPN combined with urea can improve the molluscieidal effect significantly.
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A simple and rapid expression and purification method of recombinant firefly luciferase was developed for bacteria detection. A modified luciferase gene from North American firefly Photinus pyralis was cloned into pET28a expression vector and the recombinant protein was produced in Escherichia coli BL21. The recombinant luciferase, equipped with a polyhistidine affinity tag, was purified by immobilized metal ion affinity chromatography (IMAC). The approach generated an abundant expression and an efficient purification of a recombinant luciferase with final yield 1.995mg/L of cell culture. Experiments on the recombinant luciferase also showed that the relative light units (RUL) of the enzyme were 5.8×108, and the specific activity was 2.9×1010 RLU/mg. By applying adenosine triphosphate (ATP) bioluminescence to detection of the coin bacteria using the recombinant protein, the ATP content of bacteria was 9.48×10-16mol/mL, and was identical to the bacteria counts (4500CFU/mL) in order of magnitude. Taken together, our results provided a simple and efficacious method of the preparation of recombinant luciferase, which could be applied in the determination of bacteria via ATP bioluminescence.
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According to the talented persons development project to the clinical medical profession promulgated by the university in 2005,we have been taking great effort on the reformation of the course system,the teaching content,method and the teaching form for organic chemistry experiment,and we compiled the new experimental teaching material of organic chemistry which has been published in the university.From the teaching practice of the class of clinical medical profession of 2005,we have obtained satisfatory teaching result.
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This paper combines the experience that university is in constructing center of experimental teaching.We thoroughly elaborated center of experimental teaching the guidelines of construction,develop process,and concrete conditions were introduced such as experiment resources integration,instruments management,network construction,personnel dispensation,model of managements establishment,laboratory open,and have obtained the obviously result.
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AIM To study the effects of dexamethasone on expressing monocyte chemoattractant protein-1(MCP-1 ) mRNA in the rats with pulmonary fibrosis, elaborate the molecular mechanism of dexamethasone (Dxs) in pulmonary fibrosis therapy. METHODS The model of pulmonary fibrosis was established by instilling bleomycin intratracheally. After treating with Dxsip, the levels of MCP-1 mRNA were determined by RT-PCR. The histological changes were observed and the numbers of inflammatory cells were counted in optical microscopy field. RESULTS The accumulation of inflammatory cells decreased markedly, and the symptom of pulmonary fibrosis was alleviated. Furthermore, Dxs evidently inhibited the expression of MCP-1 mRNA in lung tissues with pulmonary fibrosis. CONCLUSION The molecular mechanism of Dxs in pulmonary fibrosis therapy was associated with inhibiting the expression of MCP-1 mRNA.
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Objective To explore the effects of microwave radiation from interphone on the electrocardiogram of occupational population. Methods 165 male security personnels (working age: 0.5-3.32 a) often exposed to microwave radiation from interphone and 80 male staffs (working age: 0.5-3.58 a) without exposure to microwave radiation in a same security guard company were selected for examination of electrocardiogram in June-July,2004. Results The prevalences of sinus arrhythmia,sinus bradycardia,and the total abnormal rate of electrocardiogram in exposure group were significantly higher than those in control group(P
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AIM To study the effects of dexamethasone on expressing monocyte chemoattractant protein 1(MCP 1 ) mRNA in the rats with pulmonary fibrosis, elaborate the molecular mechanism of dexamethasone (Dxs) in pulmonary fibrosis therapy. METHODS The model of pulmonary fibrosis was established by instilling bleomycin intratracheally. After treating with Dxs ip , the levels of MCP 1 mRNA were determined by RT PCR. The histological changes were observed and the numbers of inflammatory cells were counted in optical microscopy field. RESULTS The accumulation of inflammatory cells decreased markedly, and the symptom of pulmonary fibrosis was alleviated. Furthermore, Dxs evidently inhibited the expression of MCP 1 mRNA in lung tissues with pulmonary fibrosis. CONCLUSION The molecular mechanism of Dxs in pulmonary fibrosis therapy was associated with inhibiting the expression of MCP 1 mRNA.
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To evaluate effects and mechanism of Captopril on BP of 2 -kidney, 1 -clip reno-vascular hypertensive rats ( 2K1C - RHR ), Platelets cytosolic free calcium, concentration ([Ca2+]i) and platelets aggregation (PAg) are measured. The results are as follows: [Ca2+ ]; and PAg increase significantly (P
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The number average molecular weights of five fractionated samples of pomelo pectin were determined osmometrically in aqueous solution. The values of Mn ranged from 4.74?104 to 1.83?144 for different fractions.From the data of osmotic pressure and intrinsic viscosity, the [?]-M relation obtained is[?]3.23?10-7M1.75in 0.9% NaCl solution of pH 4.83 at 37℃.