On-line high-performance liquid chromatography coupled with biochemical detection method for screening of α-glucosidase inhibitors in green tea.

An on-line high-performance liquid chromatography-biochemical detection (HPLC-BCD) method, in which compounds separated by HPLC were on-line reacted with enzyme and substrate solutions delivered by flow injection and the enzyme inhibition signal was collected by UV detection, was developed to rapidly screen α-glucosidase inhibitors from green tea extracts in this study. The chromatographic fingerprints and enzyme inhibition profiles of the different brands of green tea could be simultaneously detected by the on-line HPLC-BCD method. Enzyme inhibition profiles were detected by the UV detector at 415 nm based on the reaction of α-glucosidase and p-nitrophenyl α-d-glucopyranoside (PNPG). PNPG (1.25 mm), α-glucosidase (0.4 U/mL) and the flow rate 0.07 mL/min were applied as optimized parameters to detect α-glucosidase inhibitors in green tea. Four components in green tea showed α-glucosidase inhibition action and three of them were identified as HHDP-galloyl glucose, (-)-epigallocatechin-3-gallate and (-)-epicatechin-3-gallate by HPLC-fourier-transform mass spectrometry (HPLC-FTMS). Two brands of green tea derived from Mengding and Enshi mountainous areas might be superior to the other samples in the prevention and treatment of diabetes owing to their stronger activities of enzyme inhibitors. The proposed on-line HPLC-BCD method could be used to rapidly identify the potential enzyme inhibitors in complex matrixes.

Multiple on-line HPLC coupled with biochemical detection methods to evaluate bioactive compounds in Danshen injection.

On-line high performance liquid chromatography (HPLC) coupled with three biochemical detection (BCD) methods was applied to evaluate bioactive components in Danshen injection. On-line HPLC-photo-diode array-fluorescence detection based on the fluorogenic substrate 7-acetoxy-1-methyl quinolinium iodide, was built to search acetylcholinesterase (AChE) inhibitors in Danshen injection. On-line HPLC coupled with the scavenging assay of 1,1-diphenyl-2-picrylhydrazyl (DPPH) and 2,2'-azinobis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) free radicals was developed to screen antioxidants. The three active profiles were obviously different. Radical scavenging profiles revealed seven strong peaks in the chromatographic fingerprint possessing obvious free radical inhibition effects, while some minor peaks exhibited stronger AChE inhibition activities. The main radical scavengers and AChE inhibitors were identified by HPLC-MS. Several unknown ingredients showing strong AChE inhibition activities needed further identification except protocatechuic aldehydrate, salvianolic acid H or I and lithospermic acid. The on-line multiple on-line HPLC-BCD methods will provide powerful tools in the field of pharmacognosy for fast-track identification of interesting and/or novel bioactive compounds.

Risk factors and precautions of inpatient suicide from the perspective of nurses: A qualitative study.

The risk factors and precautions of inpatient suicide were explored. Thirty suicide victims were drawn from the adverse event reports of suicidal act during hospitalization in a general hospital from 2008 to 2014. Data were gathered from the focus group interviews of twelve nurses who had experienced inpatient suicide. The data were analyzed by using analytical technique based on grounded theory, and software QSR NVIVO8 was used to aid the collation of data. Three main themes of risk factors about inpatient suicide emerged from the analysis: individual value, social factors and environmental factors. The individual value was categorized into different groups such as sense of guilt, hopelessness and low self-esteem. Social factors included two aspects of negative life events and social support. Three themes of precautions about inpatient suicide appeared in this study: evaluation, nursing and information exchange. Evaluation was elaborated from both physical and psychological assessments. This finding extends existing work of risk factors and precautions about inpatient suicide and brings new knowledge about the reasons why inpatients commit suicide.

Diagnosis of recurrent uterine cervical cancer: PET versus PET/CT: a systematic review and meta-analysis.

OBJECTIVE: The aim of this work was to assess and compare the overall value of stand-alone FDG PET and PET/CT in diagnosing recurrent cervical cancer with a meta-analysis. METHODS: All the English published studies which addressed the use of PET whether interpreted with or without the use of CT for the diagnosis of recurrent cervical cancer were collected. Methodological quality of the included studies was evaluated. Pooled sensitivity and specificity were calculated, summary receiver operating characteristics (SROC) curve analysis was used to compare the diagnostic ability of stand-alone PET and PET/CT. RESULT: A total of 18 studies were included in this meta-analysis, with a total of 762 subjects. Pooled sensitivity and specificity of PET and PET/CT were 0.91 (95 % CI 0.87-0.94) and 0.94 (95 % CI 0.89-0.97), and 0.92 (95 % CI 0.91-0.94) and 0.84 (95 % CI 0.74-0.91), respectively. The areas under the SROC curve (AUCs) of PET and PET/CT were 0.9610 and 0.9491, respectively. There was no statistical significance between the AUC of PET and PET/CT (P > 0.05). CONCLUSION: Both PET and PET/CT have good performance in the detection of recurrent cervical cancer. However, interpreted CT images may have limited additional value on PET in detecting recurrent cervical cancer.

Identification and determination of the major triterpenes in Rhizoma Alismatis by HPLC-evaporative light scattering detection and HPLC/electrospray ionization-MS.

A simple and reliable HPLC-evaporative light scattering detection method was established for the determination of seven triterpenes in Rhizoma Alismatis, a commonly used herbal medicine. HPLC coupled with electrospray ionization-MS was applied to identify the triterpenes. The positive ion mode was used in MS detection, and the fragmentation patterns of the analytes were proposed. The quantitative method was validated for linearity, sensitivity, precision, and accuracy. All calibration curves showed good linearity (r2 > 0.9943) within the test ranges. This method showed good reproducibility with intraday and interday variations of less than 3.39 and 5.20%, respectively. The mean recoveries ranged from 96.06 to 103.5%. The proposed method was successfully applied for the quantitative analysis of the triterpenes in samples from four different habitats. The results indicated great variation of the contents of these components among the samples, and the developed assay could be considered as a suitable quality evaluation method for Rhizoma Alismatis.

Screening and identification of tyrosinase inhibitors in edible plant materials by on-line UPLC-enzyme reactor coupled with UHPLC-FTMS.

Tyrosinase plays a primary role in melanin biosynthesis and enzymatic browning of freshly cut fruits and vegetables. Herein, an on-line ultraperformance liquid chromatography diode array detector biochemical detection (UPLC-DAD-BCD) method was established to identify trace amount potent tyrosinase inhibitors and antibrowning agents in complex mixtures. The tyrosinase inhibition activities of some representative compounds were evaluated by using the established method and their chromatography-activity relationships were obtained. Then the proposed UPLC-DAD-BCD method was applied to screen tyrosinase inhibitors in edible herbal extracts and identified two tyrosinase inhibitors in green tea and three in cinnamon. The above active ingredients were determined by ultra-high-performance liquid chromatography linear ion trap/orbitrap high resolution mass spectrometry (UHPLC-FTMS). The on-line UPLC-DAD-BCD in combination with UHPLC-FTMS was confirmed to be a powerful technique to screen and elucidate the active ingredients in complex matrixes and could be applied to evaluate the integrated effects of multiple ingredients against corresponding targets.

Simultaneous screening, identification, quantitation, and activity evaluation of six acetylcholinesterase (AChE) inhibitors in Coptidis Rhizoma by online UPLC-DAD coupled with AChE biochemical detection.

Acetylcholinesterase (AChE) inhibition is a common treatment in the early stages of dementia and neurodegenerative diseases, including Alzheimer's disease (AD). Coptidis Rhizoma (CR), a traditional Chinese medicine (TCM), contains numerous isoquinoline alkaloids that substantially inhibit AChE played neuroprotective effects in the treatment of cognitive diseases. We established a method using an ultra-high-performance liquid chromatography-diode array detector coupled with AChE biochemical detection (UPLC-DAD-AChEBCD) to screen and identify AChE inhibitors (AChEIs), measure AChEIs content and activity, and evaluate the quality of CR derived from different plant species and growth year. The chromatographic fingerprint and AChEIs activity profiles of CR were simultaneously obtained by UPLC-DAD-AChEBCD, and six alkaloids including groenlandicine, coptisine, epiberberine, jatrorrhizine, berberine, and palmatine, were identified by UPLC-electrospray ionization-tandem mass spectrometry. Data analysis based on AChEIs content and total activity of 12 batches of CR indicated differences among different species and growth year. Therefore, the online method could be used to rapidly identify AChEIs in complex matrixes and screen potential agents for neurodegenerative prevention and treatment, as well as provide information for the identification and quantitation of active markers directly associated with herbal medicine quality.

Comparative study of Puerariae lobatae and Puerariae thomsonii by HPLC-diode array detection-flow injection-chemiluminescence coupled with HPLC-electrospray ionization-MS.

An on-line HPLC-diode array detection-flow injection chemiluminescence (HPLC-DAD-FICL) method was applied to estimate the difference of Puerariae lobatae and Puerariae thomsonii. Their chemical and active profiles could be obtained by HPLC-DAD-FICL in one run. Seventeen compounds in two species were tentatively identified by HPLC-electrospray ionization-MS (HPLC-ESI-MS) method. The main antioxidants were rapidly screened by active fingerprints coupled with MS data. Similarity and Hierarchical clustering analysis (HCA) were used to distinguish different samples. The results suggested that the chemical fingerprints of 16 batches of samples were similar by similarity evaluation, while HCA could discriminate the two species. The active fingerprints of Puerariae lobatae and Puerariae thomsonii were significantly different. More antioxidants were found in Puerariae lobatae than in Puerariae thomsonii. Main antioxidants, including 3'-hydroxypuerarin, genistein 8-C-glycoside-xyloside, puerarin, 6″-O-xylosylpuerarin, mirificin and daidzein in two species, may be reasonable markers for the discrimination of the two species. The integrated fingerprint based on the chemical and active characteristics may provide an objective quality evaluation for Puerariae lobatae and Puerariae thomsonii.

"Quantity-effect" research strategy for comparison of antioxidant activity and quality of Rehmanniae Radix and Rehmannia Radix Praeparata by on-line HPLC-UV-ABTS assay.

BACKGROUND: Quantitation analysis and chromatographic fingerprint of multi-components are frequently used to evaluate quality of herbal medicines but fail to reveal activity of the components. It is necessary to develop a rational approach of chromatography coupled with activity detection for quality assessment of herbal medicines. METHODS: An on-line HPLC-ultraviolet detection-2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) free radical scavenging (HPLC-UV-ABTS) method was developed to obtain the chromatographic fingerprints and ABTS+• inhibition profiles (active fingerprints) of Rehmanniae Radix (Dihuang) and Rehmannia Radix Praeparata (Shu Dihuang). Eighteen compounds showing ABTS+• inhibition activity were identified by HPLC-fourier-transform mass spectrometry (HPLC-FTMS). Verbascoside was used as a positive control to evaluate the total activities of the samples and the contribution rate of each compound. The similarities of the chromatographic and active fingerprints were estimated by the vectorial angle cosine method. RESULTS: The results showed that the HPLC-UV-ABTS method could efficiently detect antioxidant activity of the herbal medicine samples. The antioxidants were different between the two herbs and several new antioxidants were identified in Shu Dihuang. A function equation was generated in terms of the negative peak area (x) and the concentrations of verbascoside (y, µg/mL), y = 2E-07 × 4 - 8E-05 × 3 + 0.0079 × 2 + 0.5755x + 1.4754, R2 = 1. Iridoid glycosides were identified as main antioxidants and showed their higher contributions to the total activity of the samples. The total contributions of the three main active components in the Dihuang and Shu Dihuang samples to the total activity, such as echinacoside, verbascoside and an unknown compound, were 39.2-58.1% and 55.9-69.4%, respectively. The potencies of the main active components in the Shu Dihuang samples were two to ten times those in the Dihuang samples. Similarity values for S12 in the chromatographic fingerprints and S03, S12 and P03 in the active fingerprints were less than 0.9. The three batches of samples might show their different quality with the other samples. CONCLUSIONS: The results suggested that the combination of "quantity-effect" research strategy and the HPLC-UV-ABTS analysis method could comprehensively evaluate the active components and quality of Dihuang and Shu Dhuang.

The antioxidant-activity-integrated fingerprint: an advantageous tool for the evaluation of quality of herbal medicines.

The activity-integrated fingerprints of Danshen injections have been established by high-performance liquid chromatography coupled with chemiluminescence detection (HPLC-CL) according to their antioxidants activity scavenging hydrogen peroxide. The authentication and validation of the activity-integrated fingerprints were performed. Then, a data-level information fusion method was employed to capture the chemical and antioxidant information encoded in HPLC-CL fingerprints. Finally, based on the fusion results, the quality of different batches of Danshen injections samples were further evaluated by similarity measure. In comparison with conventional fingerprints, the activity-integrated fingerprints, which simultaneously contained the chemical characteristics and antioxidant activities of constitutions could comprehensively and properly reveal the quality characteristics of the Danshen injections. In conclusion, the antioxidant-activity-integrated fingerprints were suitable for quality control of Danshen injections. This study could clearly demonstrate that the activity-integrated fingerprint was a powerful and meaningful tool to comprehensively improve the quality control of complex herbal medicines.

[Classification and application principles of optical-fibre transducer].

One of the new transducers is optical-fiber transducer, which has a wideying application prospect. The present paper describes the principles, classifications and advantages of the fiber sensor, and also reports the basic features and the outline of the application of the fiber sensors in types of intensity, phase and polarization modulation.

Huachansu mediates cell death in non-Hodgkin's lymphoma by induction of caspase-3 and inhibition of MAP kinase.

Huachansu (HCS), a hot water extract of the skin glands of Bufo gargarizans (B. melanostictus), has been used extensively in the treatment of various solid tumors in Asia, particularly in China. However, its effect on the growth of malignancies of hematopoietic origin, particularly lymphomas, is limited. Here we investigated the antiproliferative effect and molecular mechanisms of HCS using non-Hodgkin's lymphoma (NHL) Raji, Ramos, and Namalwa cells and the mantle cell lymphoma cells SP53. HCS inhibited proliferation in these cell lines with an IC50 ranging from 3.1 to 25 µl/ml. At a concentration of 25 µl/ml, HCS triggered a sub-G1 arrest in Ramos cells and induced early to late apoptotic cell death. Cleaved caspase-3 was formed in a concentration-dependent manner in Ramos cells following treatment with HCS for 24 h. Intriguingly, when the Ramos cells were treated with the caspase inhibitor ZDEVD, the apoptotic activity of HCS was partially blocked. Furthermore, HCS also blocked the expression of survivin and pRB proteins in a concentration-dependent manner in Ramos cells. Mechanistically, HCS downregulated both the MAPK gene and proteins in Ramos cells. Collectively, our data suggest that HCS is effective in inducing cell death and apoptosis, in part, by activating caspase-3 activity and suppressing MAP kinase in NHL cells.

[Study on pollution for the photoelectronic material InP].

The mass spectrum analysis of crystal face (100) and (111) and the photoluminescence analysis of crystal face (100) in the photoelectronic material InP were given. The Hall coefficient, charge carrier concentration and Hall mobility were determined. Experimental results indicate that the pollution of silicon is predominant.

[Detection of DNA damage of human sperm using single cell electrophoresis].

OBJECTIVES: To detect the sperm DNA damage and to evaluate its significance in male reproductive using single cell gel electrophoresis (SCGE). METHODS: Four hundred and eighteen sperm samples were analysed using the computer assisted analysis system and SCGE. The sperms samples were divided into five grades according to the extent of the sperm nuclear DNA damage. RESULTS: 1. When the sperm density is less than 20 x 10(6)/ml, the occurence of grade II and III are increased significantly; 2. In the unmotile grade d sperm the occurence of grade I comet amounts was 5.39%, the occurence of grade II and III was remarkably increased. There was a evidently variance between the grade d and grade a + b sperm. CONCLUSIONS: SCGE can be used to detect the sperm DNA breakage and to evaluate the sperm quality and damage.

[Evaluation of radiation damage to the sperm DNA of radar operators].

OBJECTIVE: To evaluate the inflicted by radar electromagnetic radiation to the sperm DNA of radar operators. METHODS: Sperm concentration, viability, motility, sperm abnormality were determined by routine sperm analysis and sperm chromatin structure assay (SCSA) in the highly exposed group(n = 88), lowly exposed group(n = 143) and control group(n = 39). RESULTS: Sperm motility, viability of the highly exposed group reduced compared with that of the lowly exposed group and control group, while sperm abnormality increased. The COMP alpha reduction of the highly exposed group indicated that the highly exposed group had a medium fertility potential. The multifactor variable analysis showed that daily working time was a dangerous factor in sperm abnormality and abstinence time was a dangerous factor in the parameter of SCSA. CONCLUSIONS: Radar radiation inflicts damage to male reproduction system and it is important to take protective measures.

[Analysis of correlation between parameters of sperm chromatin structure assay and semen routine analysis].

OBJECTIVES: To analysis the correlation between parameters of sperm chromatin structure assay(SCSA) and semen routine analysis, and to discuss the reliable methods of semen quality evaluation. METHODS: Five hundred and eleven semen samples were detected to analyse the mutiple-parameter correlation between results of SCSA (COMP alpha t) and semen routine analysis. RESULTS: The parameters that have low-level positive correlation(r: 0.10-0.30) with denatured sperm percentage(COMP alpha t) were viscosity, ejaculation interval, abnormal sperm ratio, concentration of grade c sperm; those having low-level negative correlation(r: -0.30(-)-0.10) were VDL, VSL and VAP; those having mid-level positive correlation (r: 0.30-0.70) were sperm concentration, percentage of grade d sperm; those having mid-level negative correlation (r: -0.70(-)-0.30) were MAD, percentage of grade a sperm and survival rate. CONCLUSIONS: Flow cytometry can be used to evaluate the percentage of denatured or injured sperm rapidly, correctly and simply. The result (COMP alpha t) correlates partly with semen parameters, and it is not a conclusively parameter compared with routine semen analysis. It is important to use SCSA to evaluate productivity under the above situation.

[Study on sperm quality of the soldiers influenced by reinforced training].

OBJECTIVES: To investigate the correlation between abnormal sperm parameters and reproductive hormones and reinforced training. METHODS: Epidemic investigation were done between two different groups by determining endocrine hormone and standard clinical sperm analysis. RESULTS: Comparison between reinforced training group and non-training group showed decreased sperm velocity but no change on sperm moving form, increased teratism sperm percentage but no change on sperm number, decreased plasma testosterone but no change on LH. CONCLUSIONS: Reinforced training has correlation with weak part of sperm parameter analysis and reproductive hormones.

[The effect of overtraining on human sperm chromatin structure].

OBJECTIVE: To identify the effects of overtraining on human sperm DNA. METHODS: Molecular epidemiological investigation of 249 men from different groups (training and non-training) was carried out by using flow cytometer to detect the integrity and damage of in situ DNA of sperm nucleus, and sperm chromatin structure assay was performed. RESULTS: The average COMPalpha(t) in training group was 11.02% while that in control group was 5.90% (P < 0.01). COMPalpha(t) was significantly correlated with sperm activity (r = 0.41, P < 0.05). CONCLUSION: Overtraining could induce sperm DNA injury and affect sperm activity, thus to decrease the potentiality of reproduction.

The Spectrum-Effect integrated fingerprint of Polygonum cuspidatum based on HPLC-diode array detection-flow injection-chemiluminescence.

AIM: To establish the Spectrum-Effect integrated fingerprint of Polygonum cuspidatum to evaluate the quality of P. cuspidatum. METHODS: An on-line HPLC-DAD-flow injection chemiluminescence (FICL) method was developed to investigate the quality of P. cuspidatum from different habitats based on the established Spectrum-Effect integrated fingerprint. RESULTS: Nineteen batches of samples of P. cuspidatum were evaluated for the similarity of their chromatographic and free radical scavenging fingerprints, and the results compared. Main antioxidants were estimated by regression analysis between peak areas of thirteen compounds and their activities. Some active compounds were identified by HPLC-ESI-MS. CONSULSIONS: The results indicated that main antioxidants in P. cuspidatum could be rapidly screened by the established Spectrum-Effect integrated fingerprint based on on-line HPLC-DAD-FICL, and would be more efficient and objective method to evaluate the quality of P. cuspidatum.

Changes in cancer cell metabolism revealed by direct sample analysis with MALDI mass spectrometry.

Biomarker discovery using mass spectrometry (MS) has recently seen a significant increase in applications, mainly driven by the rapidly advancing field of metabolomics. Instrumental and data handling advancements have allowed for untargeted metabolite analyses which simultaneously interrogate multiple biochemical pathways to elucidate disease phenotypes and therapeutic mechanisms. Although most MS-based metabolomic approaches are coupled with liquid chromatography, a few recently published studies used matrix-assisted laser desorption (MALDI), allowing for rapid and direct sample analysis with minimal sample preparation. We and others have reported that prostaglandin E3 (PGE3), derived from COX-2 metabolism of the omega-3 fatty acid eicosapentaenoic acid (EPA), inhibited the proliferation of human lung, colon and pancreatic cancer cells. However, how PGE3 metabolism is regulated in cancer cells, particularly human non-small cell lung cancer (NSCLC) cells, is not fully understood. Here, we successfully used MALDI to identify differences in lipid metabolism between two human non-small-cell lung cancer (NSCLC) cell lines, A549 and H596, which could contribute to their differential response to EPA treatment. Analysis by MALDI-MS showed that the level of EPA incorporated into phospholipids in H596 cells was 4-fold higher than A549 cells. Intriguingly, H596 cells produced much less PGE3 than A549 cells even though the expression of COX-2 was similar in these two cell lines. This appears to be due to the relatively lower expression of cytosolic phospholipase A2 (cPLA2) in H596 cells than that of A549 cells. Additionally, the MALDI-MS approach was successfully used on tumor tissue extracts from a K-ras transgenic mouse model of lung cancer to enhance our understanding of the mechanism of action of EPA in the in vivo model. These results highlight the utility of combining a metabolomics workflow with MALDI-MS to identify the biomarkers that may regulate the metabolism of omega-3 fatty acids and ultimately affect their therapeutic potentials.