Analysis of full and empty ratio of EV71 virus by using capillary zone electrophoresis.

Hand, foot, and mouth disease is a serious public health problem, and the main pathogen is enterovirus 71 (EV71). Its capsid assembly mechanism including capsid protein processing has been widely studied. Full and empty capsids have different immunological efficacy. Therefore, tracking full/empty capsid ratio throughout the EV71 production process is important to ensure consistent product quality and proper dosing response. The analysis of full/empty capsid ratio of intact virus has been widely reported as well. A variety of techniques have been employed to evaluate the full/empty capsid ratios. However, there has not been a rapid, reproducible, and robust assay to determine the full/empty capsid ratios of final and in-process products. In this study, a novel assay based on capillary zone electrophoresis was established. The separation of full and empty species could be achieved within 10 min and the ratio of peak areas was used to calculate the full/empty capsid ratio directly. The results showed good reproducibility and linearity for the determination of full/empty capsid ratios.

Automated Breast Volume Scanner (ABVS)-Based Radiomic Nomogram: A Potential Tool for Reducing Unnecessary Biopsies of BI-RADS 4 Lesions.

Improving the assessment of breast imaging reporting and data system (BI-RADS) 4 lesions and reducing unnecessary biopsies are urgent clinical issues. In this prospective study, a radiomic nomogram based on the automated breast volume scanner (ABVS) was constructed to identify benign and malignant BI-RADS 4 lesions and evaluate its value in reducing unnecessary biopsies. A total of 223 histologically confirmed BI-RADS 4 lesions were enrolled and assigned to the training and validation cohorts. A radiomic score was generated from the axial, sagittal, and coronal ABVS images. Combining the radiomic score and clinical-ultrasound factors, a radiomic nomogram was developed by multivariate logistic regression analysis. The nomogram integrating the radiomic score, lesion size, and BI-RADS 4 subcategories showed good discrimination between malignant and benign BI-RADS 4 lesions in the training (AUC, 0.959) and validation (AUC, 0.925) cohorts. Moreover, 42.5% of unnecessary biopsies would be reduced by using the nomogram, but nine (4%) malignant BI-RADS 4 lesions were unfortunately missed, of which 4A (77.8%) and small-sized (<10 mm) lesions (66.7%) accounted for the majority. The ABVS radiomics nomogram may be a potential tool to reduce unnecessary biopsies of BI-RADS 4 lesions, but its ability to detect small BI-RADS 4A lesions needs to be improved.

Enhanced anticancer activity by the combination of vinpocetine and sorafenib via PI3K/AKT/GSK-3ß signaling axis in hepatocellular carcinoma cells.

Vinpocetine is widely used to treat cerebrovascular diseases. However, the effect of vinpocetine to treat hepatocellular carcinoma (HCC) has not been investigated. In this study, we revealed that vinpocetine was associated with antiproliferative activity in HCC cells, but induced cytoprotective autophagy, which restricted its antitumor activity. Autophagy inhibitors improved the antiproliferative activity of vinpocetine in HCC cells. Sorafenib is effective to treat advanced HCC, but the effect of autophagy induced by sorafenib is indistinct. We demonstrated vinpocetine plus sorafenib suppressed the cytoprotective autophagy activated by vinpocetine in HCC cells and significantly induced apoptosis and suppressed cell proliferation in HCC cells. In addition, vinpocetine plus sorafenib activates glycogen synthase kinase 3ß (GSK-3ß) and subsequently inhibits cytoprotective autophagy induced by vinpocetine in HCC cells. Meanwhile, overexpression of GSK-3ß was efficient to increase the apoptosis induced by vinpocetine plus sorafenib in HCC cells. Our study revealed that vinpocetine plus sorafenib could suppress the cytoprotective autophagy induced by vinpocetine and subsequently show synergistically anti-HCC activity via activating GSK-3ß and the combination of vinpocetine and sorafenib might reverse sorafenib resistance via the PI3K/protein kinase B/GSK-3ß signaling axis. Thus, vinpocetine may be a potential candidate for sorafenib sensitization and HCC treatment, and our results may help to elucidate more effective therapeutic options for HCC patients with sorafenib resistance.

[Rapid Start-up of a Nitrite-Dependent Methane Anaerobic Oxidation Reaction Under Static Pressure Conditions].

The present study explores the effect of static pressure on the rapid start-up of a nitrite-dependent anaerobic methane oxidation (N-DAMO) process in lab-scale sequencing batch reactors (SBR). A mixture of anaerobic sludge and deep paddy soil with a volume ratio of 1:1 was used as inoculum and the influent of the nitrite (NO2--N) concentration was gradually increased to avoid a toxicity shock. The variation of the NO2--N removal performance and corresponding microbial characteristics were analyzed to evaluate the development of the N-DAMO process. After 120 days of operation, significant N-DAMO phenomena were observed in both the control SBR (R1) with normal pressure and pressurized SBR (R2) with a static pressure of 0.3 MPa. The NO2--N removal rate (measured by NO2--N) of R2 (36.90 mg·(L·d)-1) was 24% higher than that of R1, while the average NO2--N removal rate in the first 4 h of the batch cycle in R2 (0.10 mmol·(L·h)-1) was 186% higher than that of R1. The mean sludge size of R2 was~2-fold larger than that of R1. Sludge in R2 also has a bigger specific surface area, which improves the mass transfer rate of methane and the N-DAMO performance. The specific activity of N-DAMO (measured by N/VSS) reached 0.29 mg·(g·h)-1 in the study period, which is approximately 2 times higher than that of R1. Moreover, the abundance of N-DAMO functional microbes Candidatus Methylomirabilish oxyfera (M. oxyfera) in R2 was 10-fold higher than that of R1. These results indicate that static pressure effectively accelerates the start-up of the N-DAMO process.

Metabolic responses to Lactobacillus plantarum contamination or bacteriophage treatment in Saccharomyces cerevisiae using a GC-MS-based metabolomics approach.

Bacteriophage can be used as a potential alternative agent for controlling Lactobacillus plantarum contamination during bioethanol production. However, how Saccharomyces cerevisiae respond against contaminative L. plantarum or added bacteriophage remains to be fully understood. In this study, gas chromatography-mass spectrometry and a multivariate analysis were employed to investigate the intracellular biochemical changes in S. cerevisiae cells that were elicited by L. plantarum contamination or bacteriophage treatment. The intracellular metabolite profiles originating from different groups were unique and could be distinguished with the aid of principal component analysis. Moreover, partial least-squares-discriminant analysis revealed a group classification and pairwise discrimination, and 13 differential metabolites with variable importance in the projection value greater than 1 were identified. The metabolic relevance of these compounds in the response of S. cerevisiae to L. plantarum contamination or bacteriophage treatment was discussed. Besides generating lactic acid and competing for nutrients or living space, L. plantarum contamination might also inhibit the growth of S. cerevisiae through regulating the glycolysis in S. cerevisiae. Moreover, increased concentrations of monounsaturated fatty acids secondary to bacteriophage treatment might lead to more membrane fluidity and promote the cell viability of S. cerevisiae.

Total synthesis and anti-viral activities of an extract of Radix isatidis.

Radix isatidis (Banlangen), a famous traditional Chinese medicine, has been used for thousands of years in China due to its anti-viral activity. Through our research, we inferred that the anti-viral activity of Radix isatidis depended on the water-soluble part. Among the components of this extract, the isoquinoline derivative 1 was isolated for the first time and has shown better anti-viral activity than other constituents. In this study, to solve the problem of sourcing sufficient quantities of compound 1, a total synthesis route is described, and several analogues are also evaluated for their anti-viral activities. Among them, compound 8 shown potent anti-viral activity with an IC50 value of 15.3 µg/mL. The results suggested that isoquinoline derivatives possessed potent anti-viral activity and are worthy further development.

[Study on water-soluble chemical constituents of Taraxacum mongolicum].

OBJECTIVE: To study the water-soluble chemical constituents of Taraxacum mongolicum. METHODS: The chemical constituents were isolated and purified by means of several chromatographic techniques and their structures were elucidated by spectroscopic methods. RESULTS: Nine compounds were isolated and identified as trans-p-coumaryl alcohol(1), trans-p-coumaryl aldehyde(2),p- hydroxybenzoate (3) , p-hydroxyphenyl-propionic acid (4) , 4-hydroxy-2, 6-dimethoxyphenol-1 -O-ß-D-glucopyranoside (5) , protocate- chuic aldehyde(6) ,rutin(7) ,quercetin(8) ,kaempferal-3-O-α-L-rhamnopyranosyl-( 1-6) -ß-D-glucopyranoside(9). CONCLUSION: Com pounds 1-6 are isolated from this plant for the first time.

[Chemical constituents from flower of Lonicera fragrantissima].

OBJECTIVE: To study the chemical constituents from the flower of Lonicera fragrantissima. METHODS: The chemical constituents were isolated and purified by means of several chromatographic techniques, and their structures were elucidated by spectroscopic methods. RESULTS: Nine compounds were isolated and identified as chlorogenic acid (1), caffeic acid (2), secologanoside (3), secoxyloganin(4), loganin (5), sucrose (6), myo-inositol (7), rutin (8), and chrysoeriol-7-O-ß-D-glucoside (9). CONCLUSION: Compounds 2-9 are obtained from this plant for the first time.

Role of TGF-ß1/Smads pathway in carotid artery remodeling in renovascular hypertensive rats and prevention by Enalapril and Amlodipine.

OBJECTIVE: To investigate the role of transforming growth factor-ß1 (TGF-ß1), Smad2/3 and Smad7 expressions in carotid artery remodeling in renovascular hypertensive rats, and also the therapeutic effect of Enalapril and Amlodipine. METHODS: The renovascular hypertensive rat (RHR) models with "two-kidney and one-clip" were established, including model group (n = 6), sham-operated group (n = 6), Enalapril group (10 mg/kg per day, n = 6), Amlodipine group (5 mg/kg per day, n = 6) and combination group (Amlodipine 2.5 mg/kg per day + Enalapril 5mg/kg per day, n = 6). The medication were continuous administrated for six weeks. Carotid artery morphological and structural changes in the media were observed by HE staining, Masson staining and immuno histochemical staining. Media thickness (MT), MT and lumen diameter ratio (MT/LD), and the expression levels of media α-smooth muscle actin (α-actin), proliferating cell nuclear antigen (PCNA), TGF-ß1, phosphorylated Smad2/3 (p-Smad2/3) and Smad7 in carotid arteries were measured. RESULTS: The media of carotid arteries in RHR model group was significantly thickened, the volume of smooth muscle cell was increased, and the array was in disorder; MT, MT/LD, the proliferation index of smooth muscle cell and collagen fiber area percentage of carotid arteries in the model group were significantly higher than those in the sham-operated group (P < 0.01). Compared to sham-operated group, the model group had significantly higher expressions of TGF-ß1 and p-Smad2/3 (P < 0.05) and lower Smad7 expression. Both Enalapril and Amlodipine improved smooth muscle hypertrophy and collagen deposition, reduced RHR carotid MT, MT/LD, proliferation index of smooth muscle cell, collagen fiber area percentage and the expressions of TGF-ß1 and p-Smad2/3 (P < 0.05), increased Smad7 expression (P < 0.05). Moreover, the combination treatment of Enalapril and Amlodipine had significantly better effects than single Amlodipine group (P < 0.05), but not single Enalapril group. CONCLUSIONS: TGF-ß1/Smads pathway may participate in the mechanism of carotid artery remodeling in RHR; the role of Amlodipine and Enalapril in inversing carotid artery remodeling may be related to the change of TGF-ß1/Smads pathway, the combination treatment of Amlodipine and Enalapril had better effects than single administration of Amlodipine.

[Determination of trace lead and cadmium in transgenic rice by crosslinked carboxymethyl konjac glucomannan microcolumn preconcentration combined with graphite furnace atomic absorption spectrometry].

A novel method was developed for the determination of trace lead and cadmium in transgenic brown rice based on separation and preconcentration with a micro column packed with crosslinked carboxymethyl konjac glucomannan (CCMKGM) prior to its determination by graphite furnace atomic absorption spectrometry. Variables affecting the separation and preconcentration of lead and cadmium, such as the acidity of the aqueous solution, sample flow rate and volume, and eluent concentration and volume, were optimized. Under optimized condition, detection limits of the method for the determination of trace lead and cadmium in transgenic brown rice were 0.11 and 0.002 microg x L(-1), respectively. The obtained results of lead and cadmium in the certified reference material (GBW10010, GBS1-1) were in good agreement with the certified values. The recoveries were in the range of 90%-103% and 93%-105% for detection of Pb and Cd in transgenic brown rice and the wild-type brown rice samples respectively. This study could provide technical support for determination of trace Pb and Cd in transgenic rice.

Development and validation of a high-performance liquid chromatography-tandem mass spectrometry method for the rapid simultaneous quantification of aconitine, mesaconitine, and hypaconitine in rat plasma after oral administration of Sini decoction.

A rapid, sensitive, and specific liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS-MS) method was developed and validated for simultaneous determination of aconitine (AC), mesaconitine (MA), and hypaconitine (HA), the three toxic constituents from Sini decoction (SND) in rat plasma. After the addition of citalopram as the internal standard (IS), plasma samples were basified with 100 microL 10% ammonium hydroxide, and then extracted with 1 mL ethyl acetate. Chromatographic separation was performed on a CN column (250 mm x 4.6 mm, 5 microm) with a mobile phase of methanol/40 mM ammonium acetate/formic acid (950:45:5, v/v/v) at the flow rate of 1.0 mL/min. Analytes were determined in a triple-quadrupole mass spectrometer in the selected reaction-monitoring (SRM) mode using electrospray source with positive mode. The method was validated over the concentration ranges of 0.01-10 ng/mL for AC, MA, and HA. The variation coefficients were always < 15% for both intraday and interday precision for each analyte. Mean accuracies were also within +/-15%. The method was proved to be sensitive, rapid, specific, accurate, and reproducible. It has been successfully applied to the pharmacokinetics study on rats after oral administration of SND.

Development and validation of a liquid chromatography-tandem mass spectrometry method for the determination of xanthinol in human plasma and its application in a bioequivalence study of xanthinol nicotinate tablets.

A sensitive, rapid liquid chromatographic-electrospray ionization mass spectrometric method for the determination of xanthinol in human plasma was developed and validated. Xanthinol nicotinate in plasma (0.5 mL) was pretreated with 20% trichloroacetic acid for protein precipitation. The samples were separated using a Lichrospher silica (5 microm, 250 mm x 4.6 mm i.d.). A mobile phase of methanol-water containing 0.1% formic acid (50: 50, v/v) was used isocratically eluting at a flow rate of 1 mL/min. Xanthinol and its internal standard (IS), acyclovir, were measured by electrospray ion source in positive selected reaction monitoring mode. The method demonstrated that good linearity ranged from 10.27 to 1642.8 ng/mL with r=0.9956. The limit of quantification for xanthinol in plasma was 10.27 ng/mL with good accuracy and precision. The mean plasma extraction recovery of xanthinol was in the range of 90.9-100.2%. The intra- and inter-batch variability values were less than 4.8% and 7.9% (relative standard deviation, R.S.D.), respectively. The established method has been successfully applied to a bioequivalence study of two xanthinol nicotinate tablets for 20 healthy volunteers.

[Neuroprotective effects of exogenous basic fibroblast growth factor on the hypoxic-ischemic brain damage of neonatal rats].

OBJECTIVE: To investigate the neuroprotective effect of basic fibroblast growth factor (bFGF) on neurological function after hypoxic-ischemic brain damage (HIBD) in neonatal rats. METHODS: Ninety-six HIBD models of neonatal Wistar rats were made by shearing right arteria carotis communis and then breathing 8% O(2)+92%N(2) for two hours. The models were divided into two groups randomly: the bFGF trial group and the normal saline control group. Each group had forty-eight rats. The other forty-eight neonatal Wistar rats were taken into the sham operation group. Forty rats were taken from each group and sacrificed on the 4 th, 7 th, 10 th, 17 th and 24 th days after the operation, respectively, The pathological changes in the brain were observed by optical microscope and the expressions of nestin and growth-associated protein-43 (GAP-43) in hippocampal CA1 region were examined with immunohistochemical staining and image quantitative analysis on the 4 th, 7 th, 10 th, 17 th and 24 th days after the operation. The spatial cognitive capability of other eight rats which were taken from each group respectively was evaluated by using the Morris water maze at the age of 30 days. RESULTS: (1) No brain damage was found in the sham operation group, the neurocytes were degenerative and necrotic in the control group of normal saline. The pathological manifestation of the brain damage in the bFGF trial group was milder than that of the normal saline control group. (2) Expression of nestin: The number of nestin-positive cells in hippocampal CA1 region of control group on the 4 th, 7 th, 10 th, 17 th and 24 th days after the operation was significantly increased compared with that of the sham operation group at all time points, and the numbers of nestin-positive cells in hippocampal CA1 region of the trial group were higher than those of the sham operation group and the control group (P < 0.01). (3) The expression of GAP-43 in hippocampal CA1 region of the neonatal rats reached peak on the 10th day after the operation in all the three groups. The integral optical density (IOD) of GAP-43 in hippocampal CA1 region of the control group was higher than that of the sham-operation group at all time points, and the IOD of GAP-43 in hippocampal CA1 region of the trial group was higher than those of the sham operation group and the control group at all time points (P < 0.01 for all). (4) The latency to escape platform in control group (51.75 +/- 11.27s) was longer than that in trial group (40.32 +/- 11.48s) and the sham operation group (36.58 +/- 10.83s) (P < 0.05); the frequency of passing through the platform in control group (2.34 +/- 2.42) was less than that in trial group (5.08 +/- 3.86) and the sham operation group (7.03 +/- 3.62) (P < 0.05). There was no significant difference between the trial group and the sham operation group (P > 0.05). CONCLUSIONS: (1) The expression of nestin and GAP-43 increased in hippocampal CA1 region of neonatal rats with HIBD, it may be involved in the activation of neural stem cells and the regeneration of neurocytes after HIBD. (2) The treatment with bFGF can improve the ability of learning and memory of neonatal rats with HIBD. (3) Exogenous bFGF could enhance the expression of nestin and GAP-43 in the brain of neonatal rats with HIBD, which may play an important role in restoration of neurons damaged due to hypoxia-ischemia.

Induction of apoptosis of human gastric carcinoma SGC-7901 cell line by 5, 7-dihydroxy-8-nitrochrysin in vitro.

AIM: To investigate the effect of 5, 7-dihydroxy-8-nitrochrysin (NOChR) on apoptosis of human gastric carcinoma SGC-7901 cell line. METHODS: SGC-7901 cells were cultured in vitro and the inhibitory effect of NOChR on proliferation of SGC-7901 cells was measured by using an MTT assay. NOChR-induced apoptosis rate of SGC-7901 cells was detected using flow cytometry (FCM) with PI staining. DNA ladder bands were observed by DNA agarose gel electrophoresis. The influence of NOChR on the proxisome proliferator-activated receptor-gamma (PPARgamma), Bcl-2 and Bax protein expression of SGC-7901 cells was analyzed by Western blot. RESULTS: MTT assay showed that NOChR markedly inhibited proliferation of SGC-7901 cells in a dose-dependent manner, and when IC(50) was 4.14 micromol/L, the potency of NOChR was 10 times than that of lead compound, chrysin (ChR, IC(50) was 40.56 micromol/L), and was similar to 5-fluorouracil (5-FU, IC(50) was 4.51 micromol/L). FCM with propidium iodide (PI) staining demonstrated that the apoptosis rates of SGC-7901 cells treated with 1.25, 5.00 and 20.00 micromol/L NOChR for 48 h were 9.8% +/- 0.2%, 36.8% +/- 1.9% and 45.5% +/- 3.5%, respectively, and were significantly higher when treated with 5.00 and 20.00 micromol/L NOChR than that with 20.00 micromol/L ChR (12.9% +/- 1.5%). DNA agarose gel electrophoresis showed that treatment of SGC-7901 cells with 20.00 micromol/L NOChR for 48 h resulted in typical DNA ladder bands of DNA of SGC-7901 cells, which could be eliminated by treating with 10.00 micromol/L GW9662, a blocker of PPARgamma. Western blot analysis revealed that after 24 h of treatment with 20.00 micromol/L NOChR, PPARgamma and Bax protein expression of SGC-7901 cells increased but Bcl-2 expression decreased; however, pre-incubation with 10.00 micromol/L GW9662 could efficiently antagonize and weaken the regulatory effect of 20.00 micromol/L NOChR on Bax and Bcl-2 protein expression of SGC-7901 cells.

[Effect of neferine on adriamycin-resistance of thermotolerant hepatocarcinoma cell line HepG2/thermotolerance].

BACKGROUND & OBJECTIVE: Nowadays, reversing the multidrug resistance (MDR) of thermotolerant carcinoma cells is a hot topic in tumor thermatology. This study was to investigate the adriamycin (ADR)-resistance of thermotolerant hepatocarcinoma cell line HepG2/thermotolerance and the effect of neferine (Nef) on the ADR-resistance of HepG2/thermotolerance cells. METHODS: Cell proliferation was measured by MTT assay. Cell apoptosis was detected by flow cytometry (FCM) with PI staining. The expression of Bcl-2 was measured by FCM using fluorescein isothiocyanate (FITC)-conjugated anti-bcl-2 antibodies. RESULTS: The proliferation rate and apoptosis rate of HepG2/thermotolerance cells cultured in 43 degrees C for 24 h were (89.6+/-5.4)% and (13.6+/-5.4)%, respectively; however, those of HepG2 cells were (23.9+/-3.6)% and (68.9+/-7.3)%, respectively. The 50% inhibition concentration (IC50) of ADR was 10.8 times higher for HepG2/thermotolerance cells than for HepG2 cells [(113.7+/-12.7) micromol/L vs. (10.5+/-2.3) micromol/L]. When treated with 1, 10, 100 micromol/L ADR at 37 degrees C for 24 h, the apoptosis rates of HepG2/thermotolerance cells were (9.3+/-2.6)%, (17.8+/-7.3)%, and (32.9+/-8.6)%, respectively, but those of HepG2 cells were (14.3+/-3.9)%, (38.9+/-6.8)%, and (62.7+/-5.9)%, respectively. In the presence of 10 and 40 micromol/L Nef, the IC50 of ADR for HepG2/thermotolerance cells was significantly decreased from (113.7+/-12.7) micromol/L to (63.7+/-5.6) micromol/L and (16.8+/-2.8) micromol/L, and the cell apoptosis induced by 10 micromol/L ADR was significantly increased from (17.8+/-4.3)% to (26.8+/-5.9)% and (34.9+/-8.7)%, respectively. Bcl-2 was overexpressed in HepG2/thermotolerance cells, whereas it was down-regulated when the cells were treated with 40 micromol/L Nef for 24 h. CONCLUSIONS: HepG2/thermotolerance cells are ADR-resistant. Nef may reverse the ADR-resistance of HepG2/thermotolerance cells by down-regulating Bcl-2 expression.

Rosiglitazone enhances fluorouracil-induced apoptosis of HT-29 cells by activating peroxisome proliferator-activated receptor gamma.

AIM: To examine whether and how rosiglitazone enhances apoptosis induced by fluorouracil in human colon cancer (HT-29) cells. METHODS: Human colon cancer HT-29 cells were cultured in vitro and treated with fluorouracil and/or rosiglitazone. Proliferation and growth of HT-29 cells were evaluated by MTT assay and trypan blue exclusion methods, respectively. The apoptosis of HT-29 cells was determined by acridine orange/ethidium bromide staining and flow cytometry using PI fluorescence staining. The expressions of peroxisome proliferator-activated receptor gamma (PPARgamma), Bcl-2 and Bax in HT-29 cells were analyzed by Western blot. RESULTS: Although rosiglitazone at the concentration below 30 micromol/L for 72 h exerted almost no inhibitory effect on proliferation and growth of HT-29 cells, it could significantly enhance fluorouracil-induced HT-29 cell proliferation and growth inhibition. Furthermore, 10 micromol/L rosilitazone did not induce apoptosis of HT-29 cells but dramatically enhanced fluorouracil-induced apoptosis of HT-29 cells. However, rosiglitazone did not improve apoptosis induced by fluorouracil in HT-29 cells pretreated with GW9662, a PPARgamma antagonist. Meanwhile, the expression of Bax and PPARgamma was up-regulated, while the expression of Bcl-2 was down regulated in HT-29 cells treated with rosiglitazone in a time-dependent manner. However, the effect of rosiglitazone on Bcl-2 and Bax was blocked or diminished in the presence of GW9662. CONCLUSION: Rosiglitazone enhances fluorouracil-induced apoptosis of HT-29 cells by activating PPARgamma.

[Investigation on the change in C-F stretching vibration of FTIR spectrum by 1H NMR method].

FTIR spectroscopy and 1H NMR method were used to investigate three organic compounds-1,2-di[-beta-(alpha-methyl-alpha'-p-substitutephenyl) thiophene]hexafluorocyclo pentene. After assigning all main infrared spectrum peaks, it was observed that the stretching vibration frequency(v) of C-F band in cyclopentene is rised gradually, and it was found that for compound I: V(as)C-F = 1 332.21 cm(-1), v(s) C-F) = 1 231.91 cm(-1) and delta(C-F) = 1 159.9 cm(-1); for compound II : v(as)C-F = 1 338.18 cm(-1), v(s) C-F = 1 254.31 cm(-1) and delta(C-F) = 1 179.22 cm(-1); and for compound III : v(as)C-F =1 360.99 cm(-1), v(s)C-F = 1 263.29 cm(-1) and delta(C-F) = 1 194.00 cm(-1). The reason for v change is discussed not only using infrared absorption theory qualitatively but also using 1H NMR method quantitatively. It was also found that for compound I H-1 = 7.503, H-2 = 7.484; for compound II H-1 = 6.896, H-2 = 7.465; and for compound III: H-1 = 6.728, H-2 = 7.413, proveing that the substitution group -F, -OCH2CH3, -N(CH3)2 influences the C-F stretching vibration by the conjugative effect and the inductive effect. The result is satisfactory and it shows that 1H NMR is a powerful tool for infrared spectrum analysis.

Procyanidin dimer B2 [epicatechin-(4beta-8)-epicatechin] suppresses the expression of cyclooxygenase-2 in endotoxin-treated monocytic cells.

The anti-inflammatory activity of the predominant procyanidin dimer in cocoa, dimer B2, was investigated in this study. Pretreatment of the procyanidin dimer B2 reduced COX-2 expression induced by the endotoxin lipopolysaccharide (LPS) in differentiated human monocytic cells (THP-1) in culture. To further elucidate the underlying mechanism of COX-2 inhibition by procyanidin, we examined their effects on the activation of extracellular signal-regulated protein kinase (ERK), Jun-terminal kinase (JNK), and p38 mitogen-activated protein kinase (MAPK), which are upstream enzymes known to regulate COX-2 expression in many cell types. Pretreatment with procyanidin dimer B2 decreased the activation of ERK, JNK, and p38 MAPK. In addition, procyanidin dimer B2 suppressed the NF-kappaB activation through stabilization of IkappaB proteins, suggesting that these signal-transducing enzymes could be potential targets for procyanidin dimer B2. By affecting the expression rather than the activity of COX-2, these in vitro data reported herein give further evidence on the anti-inflammatory protection by procyanidins.