Lidocaine suppresses the malignant behavior of gastric cancer cells via the c-Met/c-Src pathway.

The present study was designed to investigate the role and mechanism of action behind the action of lidocaine in gastric cancer cells. Lidocaine was tested for its potential role in affecting the viability of cells using Cell Counting Kit-8 (CCK-8) assays. It was found that there was a decreased MKN45 cell viability upon lidocaine treatment in a dose-dependent manner. Phosphorylated c-Met, phosphorylated c-Src, c-Met and c-Src levels were detected using western blotting following lidocaine or hepatocyte growth factor (HGF) intervention. It was found that the phosphorylation levels of c-Met and c-Src were markedly reduced by lidocaine treatment, with this effect being further relieved by the addition of HGF. Subsequently, whether lidocaine repressed the malignant biological properties of gastric cancer cells through the c-Met/c-Src axis was further investigated through the detection of epithelial-mesenchymal transition markers (N-caderin and vimentin), wound healing and transwell assay analysis. In addition, cell apoptosis and the levels of apoptosis-related proteins were determined using TUNEL and western blot assays, respectively. The results demonstrated that the malignant behavior of cells were notably repressed upon lidocaine treatment, but the addition of HGF markedly reversed these effects, indicating that the effects of lidocaine on supressing the malignant behaviour of cells could be mediated through the c-Met/c-Src axis. Subsequently, whether lidocaine affected the sensitivity of cells to cisplatin or 5-FU was analyzed using a CCK-8 assay. Enhanced sensitivity of cells to cisplatin or 5-FU was observed when treated in combination with lidocaine. The present study concluded that the involvement of the c-Met/c-Src pathway in the biological behaviour of MKN45 cells was mediated by lidocaine. Therefore, lidocaine may have the potential to suppress the malignant behaviour and proliferation of gastric cancer cells.

[Identification of Efficient Strain Applied to Mining Rehabilitation and Its Rock Corrosion Mechanism:Based on Boosted Regression Tree Analysis].

The soil and water loss problem in rock mining areas is an extremely serious problem,and microbes play significant roles in ecological restoration of those areas.In this study,directive screening was used to explore the efficiency of microbe-mediated habitat restoration and the underlying mechanisms.A bacterial strain NLX-4,which was then identified as Pseudomonas protegens according to its 16S rRNA gene sequence,was screened out as an efficient silicate dissolution bacterium with the ability to secrete siderophore and indole-3-acetic acid (IAA).Moreover,pH value,element (K,Al,Si) release,organic acid content,amino acid concentration,polysaccharide content,and rock particle diameter variation in culture medium were analyzed to explore the ability of P.protegens NLX-4 to promote dolomite dissolution under controlled experimental conditions.These results showed that P.protegens NLX-4 could play a positive role in dolomite dissolution by producing tartaric acid (>777 mg·L-1) and polysaccharides (>8.21g·L-1).Therefore,P.protegens NLX-4 is an efficient microbial resource that can be used in rehabilitation of abandoned mines and has great application potential.

[Authentication and adulteration analysis of sesame oil by FTIR spectroscopy].

It's common in edible oil market that adulterating low price oils in high price oils. Sesame oil was often adulterated because of its high quality and price, so the authentication and adulteration of sesame oil were qualitatively and quantitatively analyzed by Fourier transform infrared (FTIR) spectroscopy combined with chemometrics. Firstly, FTIR spectra of sesame oil, soybean oil, and sunflower seed oil in 4,000-650 cm(-1) were analyzed. It was very difficult to detect the difference among the spectra of above edible oils, because they are all mixtures of triglyceride fatty acids and have similar spectra. However, the FTIR data of edible oils in the fingerprint region of 1,800-650 cm(-1) differed slightly because their fatty acid compositions are different, so the data could be classified and recognized by chemometric methods. The authenticity model of sesame oil was built by principal component analysis (PCA) and soft independent modeling of class analogy (SIMCA). The recognition rate was 100%, and the built model was satisfactory. The classification limits of both soybean oil and sunflower seed oil adulterated in sesame oil were 10%, with the chemometric treatments of standard normal variation (SNV), partial least square (PLS) and PCA. In addition, the FTIR data processed by PCA and PLS were used to establish an analysis model of binary system of sesame oil mixed with soybean oil or sunflower oil, the prediction values had good corresponding relationship with true values, and the relative errors of prediction were between -6.87% and 8.07%, which means the quantitative model was practical. This method is very convenient and rapid after the models have been built, and can be used for rapid detection of authenticity and adulteration of sesame oil. The method is also practical and suitable for the daily analysis of large amount of samples.

[Raman spectroscopy combined with pattern recognition methods for rapid identification of crude soybean oil adulteration].

In the present paper, a non-destructive, simple and rapid analytical method was proposed based on Raman spectroscopy (Raman) combined with principal component analysis (PCA) and support vector machine (SVM) as pattern recognition methods for adulteration of crude soybean oil (CSO). Based on fingerprint characteristics of Raman, the spectra of 28 CSOs, 46 refined edible oils (REOs) and 110 adulterated oil samples were analyzed and used for discrimination model establishment. The preprocessing methods include choosing spectral band of 780-1,800 cm(-1), Y-axis intensity correction, baseline correction and normalization in succession. After those series of spectral pretreatment, PCA was usually employed for extracting characteristic variables of all Raman spectral data and 7 principal components which were the highest contributions of all data were used as var- iables for SVM model. The SVM discrimination model was established by randomly picking 20 CSOs and 95 adulterated oils as calibration set, and 8 CSOs and 35 adulterated oils as validation set. There were 4 kinds of kernel function algorithm (linear, polynomial, RBF, sigmoid) respectively used for establishing SVM models and grid-search for optimization of parameters of all the SVM models. The classification results of 4 models were compared by their discrimination performances and the optimal SVM model was based on linear kernel classification algorithm with 100% accuracy rate of calibration set recognition, a zero misjudgment rate and the lowest detection limit of 2.5%. The above results showed that Raman combined PCA-SVM could discriminate CSO adulteration with refined edible oils. Since Raman spectroscopy is simple, rapid, non-destructive, environment friendly, and suitable for field testing, it will provide an alternative method for edible oil adulteration analysis.

Oxidative damage in various tissues of rats exposed to radon.

Oxidative damage can be induced by many environmental stressors. 8-Hydroxydeoxyguanosine (8-OHdG) has been used as a biomarker of oxidative DNA damage in both in vitro and in vivo studies. In the present study, Wistar rats were exposed to radon gas at a concentration of 100,000Bq/m(3) for 12 h/d for 30, 60, and 120 d, equivalent to cumulative doses of 60, 120, and 240 working level months (WLM), respectively. Changes in levels of 8-OHdG, reactive oxygen species (ROS), and total antioxidant (T-AOC), as well as expressions of some DNA repair enzymes such as 8-oxoguanine DNA glycosylase (OGG1) and MutT homolog 1 (oxidized purine nucleoside triphosphatase, MTH1), were determined in rat urine, peripheral blood lymphocytes, and lung after exposure to radon. The results revealed an increase in 8-OHdG and ROS levels, a decrease in T-AOC levels, and reduced OGG1 and MTH1 expression levels. The elevated amount of 8-OHdG in urine or lymphocytes was positively correlated with the cumulative exposure dose, whereas OGG1 and MHT1 expression levels in lung were inversely correlated with cumulative exposure dose. These findings indicate that oxidative damage induced by radon may be involved in radon-induced carcinogenesis.

[Rapid determination of Sudan dyes adulterated in natural paprika red by FTIR].

How to rapidly determine synthesized pigments or dyes adulterated in natural pigments is a difficult problem for food analysts. Natural paprika red is widely used in foods because of its coloration and pharmaceutical effect, but it is sometimes adulterated with Sudan dyes for its poor stability under light or heat treatment. In the present paper, the macro-fingerprint characteristic of infrared spectroscopy was utilized to identify Sudan dyes adulterated in paprika red. The strong peaks at 1 621, 1 500 and 751 cm(-1) in FTIR spectra and at the fingerprint region of 753, 684 and 496 cm(-1) in the secondary deriative FTIR spectra were remarkable characteristics for Sudan dyes adulterated in paprika red, of which the limit of determination was about 1%.

[Study on the processing of leech by FTIR and 2D-IR correlation spectroscopy].

The chemical differences of traditional Chinese medicine leech before and after processing were analyzed by FTIR and two-dimensional correlation infrared (2D-IR) spectroscopy. The result showed that the leech was high in protein, with characteristic peaks of amide I, II bands. Comparing the IR spectra of samples, the primary difference was that the characteristic peak of fresh leech was at 1 543 cm(-1), while that of crude and processed leech was at 1 535 cm(-1). A 2D-IR spectrum with heating perturbation was used to track the processing dynamics of leech In the 2D-IR correlation spectra, fresh leech exhibited stronger automatic peaks of the amide I and II bands than that of processed leech, which indicates that the protein components of the fresh leech were more sensitive to heat perturbation than the processed one. Moreover, the result of FTIR and 2D-IR correlation spectra validated that the 3-dimensional structure of protein was damaged and hydrogen bonds were broken after processing, which resulted in the inactivation of protein. The fatty acids and cholesterol components of leech were also oxidized in this process.

[Mechanism study of adaptive response in high background radiation area of Yangjiang in China].

OBJECTIVE: To study the adaptive response mechanisms in high background radiation area (HBRA) among Yangjiang local people through gene and protein expression of receptor for advanced glycation end products (RAGE) and S100A6 in peripheral blood and sputum in inhabitants of HBRA. METHODS: A total of 53 male inhabitants were selected from HBRA in Yangjiang as the exposure group, while 53 male inhabitants were selected from Enping (control area, CA)as the control group. The content of RAGE and S100A6 gene and protein were detected by RT-PCR and Western blotting assay. Thermo luminescent dosemeter(TLD) assay was used to measure the outside dose and estimate the effective dose. RESULTS: The effective dose in CA and HBRA was respectively 1.95 mSv and 6.24 mSv, which was 3 fold difference. Compared with CA, RAGE and S100A6 expression were significantly reduced in both gene and protein level in HBRA. The relative median mRNA expression of RAGE and S100A6 in peripheral blood were respectively 0.28, 1.06 and 0.16, 0.79 in CA and HBRA group, there was significance (with analysis Z values of -2.587 and -2.328 respectively, P < 0.05) with Wilcoxon rank test. For the protein of sputum, the relative median expression were respectively 2.98, 2.25 and 0.53, 0.47 with significant difference (with analysis Z values of -2.201 and -2.366 respectively, P < 0.05) by Wilcoxon rank test. CONCLUSION: The low expression of RAGE and S100A6 in HBRA group might be correlated with the adaptive response and the low mortality of cancer in HBRA.

[Identification of musk by Fourier transform infrared spectroscopy].

Standard nature musk and false nature musk were identified by Fourier transform infrared spectroscopy (FTIR) combined with derivative spectra in the present article. The main characteristic absorption peaks of musk represented the key components such as macrocyclic ketone (2923, 2851 cm(-1)), polypeptides (1655, 1546 cm(-1)) and steroids (1400, 1038 cm(-1)) respectively. Ensuing from the researches artificial musk and natural musk resemble the exemplar of market nature musk. Particularly, the infrared spectra of samples of natural musk and artificial musk are more similar and hard to be distinguished. In virtue of derivative spectra analysis, the resolution of spectra is advanced; several kinds of standard musk have been differentiated directly and effectively. This method was fast, sensitive, direct and nondestructive, and could be used to identify the source of rare materia medica and distinguish the counterfeits of musk successfully.

[Study on esterified modification of anthocyanins by FTIR].

Anthocyanins are relatively abundant in vegetables and fruits, which have potential positive health effects. The role of anthocyanins as food coloring agents becomes very important because they can provide attractive bright color of many food products. Nevertheless, the instability of natural anthocyanins was a big obstacle for its usage in food as colorants. The stability of the red radish anthocyanins is significantly improved by modified esterification of the colorant. Usually, the red radish anthocyanins was composed of several components of similar structures. The major methods for determining the structures of anthocyanin colorants involve chromatographic techniques such as TLC, HPLC and HPLC-MS, which are very useful in separation and identification of the components of anthocyanins However, compared to the spectroscopic method, the chromatographic methods are usually complicated and time-consuming during separation and analysis. In the present paper, the authors seek to establish a new, rapid and economic method for the analysis of structural change before and after esterified modification of anthcyanins in view of unique macro-fingerprint characteristics of infrared spectroscopy, which could reflect the whole change of complicated mixture system. The anthocyanins from red radish was esterification-modified by reacting with succinic anhydride, and the natural and modified anthocyanins were detected by FTIR The results showed that carbonyl of succinic anhydride was connected with the hydroxyl in glucosyl rings of anthocyanins to form new esterified anthocyanins, which are more stable than the natural one and present attractive bright color as usual.

[Determination of trace BrO3- and Br- in Chinese patent drugs, waters and beverages by ion chromatography coupled with inductively coupled plasma mass spectrometry].

In the present study, a method for determination of BrO3- and Br- using ion chromatograghy coupling with inductively coupled plasma mass spectrometry (IC-ICP-MS) was developed. BrO3- and Br- were separated on a Hamilton PRP X-100 (150 x 4.1 mm, 10 microm) column guarding with a On-guard II RP (4 x 50 mm) column with 10 mmol x L(-1) NH4NO3 eluent solution at 2 mL x min(-1). By using 200 microL sample loop, the detection limits for BrO3- and Br were 0.35 and 0.36 microg x L(-1), respectively. BrO3- had good linearity in the range 4.8-160.0 ng x L(-1). The standard solution linear equation was y = 250.31x-45.43, and R2 = 0.9999. Br had good linearity in the range 4.2-140.0 ng x L(-1). The standard solution linear equation was y = 186.84x-127.10, and R2 = 0.9994. Recoveries of spiked samples were 98.9%-109.5% and 97.4%-106.1%, respectively. The samples included various kinds of Chinese patent drugs, waters, and beverages. Among them, 14 bottled drinking waters and 2 kinds of tap waters were found containing BrO-, while other samples were under BrO3- detection limit. Br- was detected in all samples.

[Study on compatibility of Coptidis-Evodia couples by FTIR spectroscopy].

Combining unique macro-fingerprint characteristics of infrared spectroscopy (IR) with the overall concept of traditional Chinese medicine (TCM) theory, the authors seek to establish a new, rapid and economic method for research on Chinese herb preparation (CHP), which could reflect the whole and integration concept of TCM theory and also be widely applied in TCM research. In the present paper, four coptidis-evodia couples, e.g. Zuo-jin-wan (coptidis : evodia = 6 : 1), Gan-lu-san (coptidis : evodia = 2 : 1), Zhu-yu-wan (coptidis : evodia = 1 : 1) and Fan-zuo-jin (coptidis : evodia = 1 : 6), were extracted by boiling water and 80% alcohol, and the FTIR spectra of those CHPs were compared and analyzed in detail. The characteristic absorbance bands of coptidis and evodia were assigned in the FTIR spectra of the four coptidis-evodia couples. The results showed that rhizoma coptidis owns absolute superiority in both extracts of Zuo-jin-wan and Gan-lu-san since the proportions of rhizoma coptidis in the two couples are higher than those of fructus evodiae. With regard to Zhu-yu-wan, the proportions of two ingredients are the same. However, the contribution rate of coptidis is higher than that of evodiae in water extract of Zhu-yu-wan. In addition, coptidis also shows higher contribution rate than that of evodia in alcohol extract of Fan-zuo-jin. The spectroscopic technique described here is a simple and rapid analytical technique, which could provide valuable chemical information about the whole coptidis-evodia CHPs.

[The effect of nitric oxide inhalation on the expression of COX-2 and toll-like receptor 4 in rat acute lung injury induced by lipopolysaccharide].

OBJECTIVE: To study the effect of nitric oxide (NO) on the expression of cyclo-oxygenase 2 (COX-2) and toll-like receptor 4 (TLR4) in a rat model of acute lung injury (ALI) induced by lipopolysaccharide (LPS). METHODS: Twenty-four Sprague-Dawley rats were randomly divided into 4 groups. Group N: normal control group; Group L: ALI model by LPS intratracheal instillation; Group Za: ALI model+inhaled NO 20x10(-6) mg/L; Group Zb: ALI model+inhaled NO 100x10(-6) mg/L. Lung morphology was studied and COX-2 was detected by immunohistochemistry (IHC) while TLR4 by fluorescent quantitative PCR (FQ-PCR). RESULTS: Immunohistochemistry and FQ-PCR showed that COX-2 (2.8+/-0.8) and TLR4 (2.1+/-0.7) were detected in the respiratory tract of the normal control rats. In Group L, the expression of COX-2 (6.5+/-2.8) and TLR4 (44.9+/-11.3) was increased in the main bronchus and bronchioles, compared to the normal controls (t=3.003, 10.480, both P<0.01). In Group Zb, the expression of COX-2 (5.0+/-2.0) and TLR4 (16.2+/-3.8) were decreased as compared to Group L, but only the level of TLR4 showed statistical difference (t=7.030, P<0.001). CONCLUSIONS: COX-2 and TLR4 distributed widely in the respiratory tract of the rats. LPS increased the expression of COX-2 and TLR4. Low dose of nitric oxide (20x10(-6) mg/L) inhalation reduced the bronchiolar expression of COX-2 and TLR4 induced by LPS.

[Study on traditional Chinese medicine extracts of rhizoma coptidis by FTIR spectroscopy].

In the present paper, we applied Fourier transform infrared spectroscopy (FTIR) and second derivative infrared spectroscopy to establish a rapid and efficient analytical method for a preliminary quality control of medicinal herb extracts. The commonly used Chinese herb medicine, Rhizoma coptidis, was taken as an example. The main component-berberine in rhizoma coptidis was analyzed as target compound. The results indicated that the IR spectra of rhizoma coptidis herb, its water and alcohol extracts, exhibit their macroscopic fingerprint characters with fine repeatability, and the corresponding secondary derivative spectra further confirmed the spectral characteristics of the original IR spectra by enhancing spectral resolution. The typical and strongest absorption band of IR spectrum of berberine is at 1,505 cm(-1), which is assigned to aromatic skeleton vibration band, and its intensities in spectra of rhizoma coptidis herb and its different extracts become the main identification standard of the contents of berberine in those samples. In addition, comparing the IR spectra of the medicinal herb and its extracts, it was shown that the main bioactive components, alkaloids in rhizoma coptidis herb, are enriched by extraction, and the contents of berberine in alcohol extracts of rhizoma coptidis are higher than those in water extracts. The spectroscopic technique described is a simple and rapid analytical technique, which could provide valuable information about chemical constituents of medicinal extracts for guiding further chromatographic analysis and separation improvement.

Up-regulation of RAGE and S100A6 in rats exposed to cigarette smoke.

Cigarette smoke has been widely investigated in terms of epidemiology and pathological endpoints in relation to human lung diseases and animal study. In this study we exposed Wistar rats to cigarette smoke at concentrations of 20% and 60% to explore potential molecular mechanisms at the protein level. Exposures were conducted twice a day, 5 days a week for 43 weeks. As a major metabolite of nicotine in cigarette, cotinine level in rat urine was determined by HPLC-MS. A dose-dependent analysis indicated that cotinine may be used as an exposure marker of cigarette smoke. Expression of receptor for advanced glycation endproducts (RAGE), an immunoglobulin super family that triggers the intracellular signal cascade reaction leading to inflammation and its ligand S100A6 (calgranulin) in bronchial epithelial cells and lung tissues of rats, were found to be positive correlated with cotinine levels, indicating that RAGE and S100A6 may be attributable to inflammation and oxidative damage caused by cigarette smoke.

Fourier transform mid-infrared (MIR) and near-infrared (NIR) spectroscopy for rapid quality assessment of Chinese medicine preparation Honghua Oil.

Honghua Oil (HHO), a traditional Chinese medicine (TCM) oil preparation, is a mixture of several plant essential oils. In this text, the extended ranges of Fourier transform mid-infrared (FT-MIR) and near infrared (FT-NIR) were recorded for 48 commercially available HHOs of different batches from nine manufacturers. The qualitative and quantitative analysis of three marker components, alpha-pinene, methyl salicylate and eugenol, in different HHO products were performed rapidly by the two vibrational spectroscopic methods, i.e. MIR with horizontal attenuated total reflection (HATR) accessory and NIR with direct sampling technique, followed by partial least squares (PLS) regression treatment of the set of spectra obtained. The results indicated that it was successful to identify alpha-pinene, methyl salicylate and eugenol in all of the samples by simple inspection of the MIR-HATR spectra. Both PLS models established with MIR-HATR and NIR spectral data using gas chromatography (GC) peak areas as calibration reference showed a good linear correlation for each of all three target substances in HHO samples. The above spectroscopic techniques may be the promising methods for the rapid quality assessment/quality control (QA/QC) of TCM oil preparations.

Comparative study on major bioactive components in natural, artificial and in-vitro cultured Calculus Bovis.

Major bioactive components in various Calculus Bovis, including natural, artificial and in-vitro cultured Calculus Bovis, were comparatively studied. An approach of high-performance liquid chromatography coupled with ultraviolet and evaporative light scattering detections (HPLC/UV/ELSD) was established to simultaneously determinate six bioactive components thereof, including five bile acids (cholic acid, deoxycholic acid, ursodeoxycholic, chenodeoxycholic acid, hyodeoxycholic acid) and bilirubin. ELSD and UV detector were applied to detect bile acids and bilirubin respectively. The assay was performed on a C(18) column with water-acetonitrile gradient elution and the investigated constituents were authenticated by comparing retention times and mass spectra with those of reference compounds. The proposed method was applied to analyze twenty-one Calculus Bovis extraction samples, and produced data with acceptable linearity, precision, repeatability and accuracy. The result indicated the variations among Calculus Bovis samples under different developmental conditions. Artificial and in-vitro cultured Calculus Bovis, especially in-vitro cultured ones, which contain total bioactive constituents no less than natural products and have the best batch-to-batch uniformity, suffice to be used as substitutes of natural Calculus Bovis.

[Establishment of a novel preliminary screening model for bioactive parts of folium of Isatis indigotica based on biothermodynamics].

OBJECTIVE: To apply a novel method for screening the bioactive parts and substances of traditional Chinese medicines (TCM) based on biothermodynamic theories. METHOD: The power-time curves of Escherchia coli proliferation at different concentrations of decoction and its five bioactive parts of folium of Isatis indigotica (FII) were determined by TAM Air Isothermal Calorimeter. The growth rate constants of promotive and inhibitory actions were calculated. Moreover, the potential pharmacological action of FII was analyzed from the point of view of TCM theory. RESULT: The growth rate constant k1 and Pm of the studied bacterium in the presence of the decoction of FII were concentration-dependant. The k1 and Pm increased with the mass increase of the decoction and its water-extract of FII. However, the value of k1 and Pm decreased with the increasing concentrations of its other four organic solvent parts. The experimental results revealed that the sequence of inhibitory activity of different extracts was n-BuOH extract > EtOAc extract > chloroform extract > PE extract. CONCLUSION: The decoction and its water-extract of FII stimulated the metabolism of E. coli. While the other four organic solvent extracts of the total decoction inhibited the growth of tested bacterium. The microcalorimetric method has many virtues such as extensive applicability, convenience, etc, and provides a new and useful means for primary screening the bioactive parts and substances of TCM.

[Phytochemical and pharmacological research progress in Leontopodium medicinal plants].

This paper reviewed the phytochemical and pharmacological research progress in Leontopodium medicinal plants, including the resource distribution, the chemical constitutes, the pharmacological activities and clinical application. The review has provided some information for the study and development of Leontopodium medicinal plants in future.