Inhibition of Liver Cancer HepG2 Cell Proliferation by Enzymatically Prepared Low-molecular Citrus Pectin.

BACKGROUND: Low-molecular citrus pectin (LCP) is a pectin polysaccharide with low molec-ular weight, low degree of crux, and no branching. It is obtained by degrading natural citrus pectin (CP) through physical, chemical and enzymatic methods. LCP has received considerable attention in recent years due to its potential applications in the medical and biological fields. METHODS: In our previous study, LCP was prepared from CP by using recombinant Bacillus subtilis pectate lyase B. Monosaccharide comparative analysis revealed that the galacturonic acid content of LCP was higher than that of CP. The cell viability effect of LCP was elucidated by using HepG2 cells and the Cell Counting Kit-8 (CCK-8) assay. Terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL) staining, Annexin V-FITC/PI staining, and flow cytometer propidium iodide stain-ing were performed to detect the effects of LCP on apoptosis and cell cycle arrest in HepG2 cells. Mi-tochondrial membrane potential (MMP) was observed through 5,5',6,6'-tetrachloro-1,1',3,3'-tetraethyl-imidacarbocyanine assay. RESULTS & DISCUSSION: The Mw of the prepared LCP was 7.6 kDa, which was significantly lower than that of CP (140 kDa). Cell viability decreased with the increase in the concentration of LCP. The half-inhibitory concentration of 1.46 ± 0.02 mg/mL was determined. Treatment with 1.6 mg/mL LCP in-duced the apoptosis of HepG2 cells with the inhibition rate of 83.10% ± 4.72%, and the cell cycle was arrested in the S phase. Furthermore, the MMP of HepG2 cells decreased with the increase in LCP concentration. CONCLUSION: The enzymatically prepared LCP could inhibit the proliferation of HepG2 cells. This study provided a partial experimental basis and reference for LCP to become a potential functional food for anti-liver cancer.

[Lingnan fire needling combined with 0.1% sodium hyaluronate eye drops in the treatment of xerophthalmia: a randomized controlled trial].

OBJECTIVE: To observe the effect of Lingnan fire needling combined with artificial tears in the treatment of xerophthalmia. METHODS: A total of 86 xerophthalmia patients were equally and randomly divided into treatment group and control group. The patients of both groups were received treatment with 0.1% sodium hyaluronate eye drops in their eyes 3 times a day, one drop in each eye. In addition, the patients of treatment group also treated by Lingnan fire needling on bilateral Shaoze (SI1), Neichengqi and beside lacrimal puncta once a week. The treatment was conducted for 4 consecutive weeks. Before and after 4 weeks of treatment, the clinical efficacy, visual acuity, intraocular pressure, ocular symptom score, OSDI score, fluorescence staining （FL） score, schemer I, tear menisci height, tear film break-up time (BUT) and eye redness index were recorded and evaluated. RESULTS: After the treatment, self-comparison showed that the symptom score, OSDI score, FL score and eye redness index were significantly decreased (P<0.05), and BUT was notably increased in both groups (P<0.05) in comparison with their own pre-treatment. The tear menisci height in the treatment group was higher than that before the treatment (P<0.05). Comparison between the two groups showed that the symptom score, OSDI score and eye redness index were obviously lower in the treatment group than in the control group (P<0.05), whereas the BUT and tear menisci height were evidently higher (P<0.05). The total effective rate of the treatment group was 84.88% (73/86), better than 76.74% (66/86) of the control group (P<0.05). CONCLUSION: Lingnan fire needling combined with 0.1% sodium hyaluronate eye drops is more effective than simple sodium hyaluronate eye drops for xerophthalmia patients, with significant curative effect and no adverse reactions.

Effects of dietary inclusion levels of palm oil on growth performance, antioxidative status and serum cytokines of broiler chickens.

This study was conducted to investigate the effects of dietary palm oil inclusion levels on growth performance, serum antioxidative status and cytokines in Sanhuang broiler chickens. A total of 208 one-day-old female Sanhuang chickens were randomly allocated to four treatment groups with four replicates per group and of 13 birds each for a 42-day feeding trial. Dietary treatments were basal diet without palm oil (control) and diets containing 2%, 4% or 6% palm oil. During days 22 to 42 and the overall experimental period, chickens fed diets supplemented with 4% and 6% palm oil had a lower average daily feed intake (ADFI) (p = 0.019) and feed to gain ratio (F/G) (p = 0.031). Average daily gain (ADG) tended to be higher (p = 0.089) for palm oil inclusion treatments compared with the control treatment from days 1 to 42. Dietary 2% and 4% palm oil treatments significantly lowed serum malondialdehyde concentration on day 21 when compared to the control and 6% palm oil treatments (p = 0.027). Dietary 6% palm oil treatment significantly reduced the catalase (CAT) activity (p = 0.018) and total antioxidant capacity (T-AOC) (p = 0.027) in serum in comparison with the control treatment on day 21. No significant differences were observed in superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) activities in serum among dietary treatments on day 21. There were no significant differences in T-AOC, CAT, SOD and GSH-Px activities among dietary treatments on day 42. Dietary inclusion of 4% and 6% palm oil significantly increased (p = 0.01) serum interleukin-1ß (IL-1ß) content, but improved transforming growth factor-ß (TGF-ß) content in serum on both days 21 and 42 (p = 0.032 and 0.022 respectively) as well as IL-10 content on day 42 (p = 0.022) when compared with the control treatment. Furthermore, dietary inclusion of 6% palm oil significantly reduced tumour necrosis factor-α (TNF-α) content on days 21 and 42 (p = 0.021 and 0.001 respectively) compared with the control treatment. These results suggest that dietary inclusion of 4% and 6% palm oil could improve the growth performance, modulate the balance between pro- and anti-inflammatory cytokines, and show limited effect on antioxidative status in Sanhuang broiler chickens.

[Establishment and Application of Model for Determining Oil Content of Cottonseed Using Near Infrared Spectroscopy].

Cotton is one of the important oil crops, and it is great significance for screening and identification of breeding materials to establish a method of the rapid, nondestructive testing of cotton seed oil content. In this study, near-infrared diffuse reflection spectroscopy of 118 high and low oil materials were adopted to establish models for fast nondestructive determining oil content of cottonseed using near infrared spectroscopy (NIR). One hundred and six cottonseed samples as calibration set that covered the range of seed oil content for upland cotton were used in this experiment. The spectral data of cottonseed were processed using the first derivative and multiplicative scatter correction (MSC). The correction NIR model of oil content was built based on partial least squares (PLS) method with the spectral regions 5 446-8 848 cm(-1) and main components (5). The determination coefficient (R2) of calibration model was 0.975, standard error of calibration (SEC) was 0.67. The authors test the model's actual ability to predict using external validation set. The correlation coefficient (r) of predicted values and the chemistry value was 0.978, the range of prediction error was 0.1% - 1.7%. The model established has good predictability. The oil content of 784 breeding stocks were predicted by NIR model, statistical analysis of predictable. results elucidated that the NIR model of oil content developed can be well applied to selective breeding and oil related study in cotton.

PPARγ-dependent pathway in the growth-inhibitory effects of K562 cells by carotenoids in combination with rosiglitazone.

BACKGROUND: Carotenoids have been found to play roles in the prevention and therapy of some cancers which PPARγ was also discovered to be involved in. The present studies were directed to determine the inhibitory effects of carotenoids in combination with rosiglitazone, a synthetic PPARγ agonist, on K562 cell proliferation and elucidate the contribution of PPARγ-dependent pathway to cell proliferation suppression. METHODS: The effects of carotenoid and rosiglitazone combination on K562 cell proliferation were evaluated by trypan blue dye exclusion assay and MTT assay. When PPARγ has been inhibited by GW9662 and siRNA, cycle-related regulator expression in K562 cells treated with carotenoid and rosiglitazone combination was analyzed by Western blotting. RESULTS: Rosiglitazone inhibited K562 cell proliferation and augmented the inhibitory effects of carotenoids on the cell proliferation greatly. Specific PPARγ inhibition attenuated the cell growth suppression induced by carotenoid and rosiglitazone combination. GW9662 pre-treatment attenuated the enhanced up-regulation of PPARγ expression caused by the combination treatment. Moreover, GW9662 and PPARγ siRNA also significantly attenuated the up-regulation of p21 and down-regulation of cyclin D1 caused by carotenoids and rosiglitazone. CONCLUSIONS: PPARγ signaling pathway, via stimulating p21 and inhibiting cyclin D1, may play an important role in the anti-proliferative effects of carotenoid and rosiglitazone combination on K562 cells. GENERAL SIGNIFICANCE: Carotenoids in combination with rosiglitazone are hopeful to provide attractive dietary or supplementation-based and pharmaceutical strategies to treat cancer diseases.

[Effects of site conditions and tree age on Robinia pseudoacacia and Populus simonii leaf hydraulic traits and drought resistance].

In order to elucidate the hydro-physiological mechanisms of the formation of 'dwarf and aged' trees, two typical tree species Robinia pseudoacacia and Populus simonii on the Loess Plateau were chosen as the test objects to study their leaf hydraulic traits and drought resistance under effects of site conditions (gully bottom tableland and inter-gully slopeland) and tree age. For both R. pseudoacacia and P. simonii, the trees growing on the wet tableland had higher leaf maximum hydraulic conductance (Kmax) and hydraulic vulnerability (P50), and the Kmax decreased with tree age but the P50 had less difference, as compared with the trees growing on the dry slopeland. The trees growing on the tableland also had higher leaf epidermal conductance and PV curve parameters (including relative water content at turgor pressure loss point RWC(tlp), water potential at turgor pressure loss point psi(tlp), and osmotic potential at saturated water content psi(sat), and the leaf epidermal conductance decreased significantly with tree age whereas the PV curve parameters had definite decrease. The Kmax was significantly positively correlated with psi(tlp), and the P50 had definite correlations with PV curve parameters, suggesting that there existed a tradeoff relationship between Kmax and drought resistance, and P50 was an important drought-resistant trait for the two tree species.

[Analysis method of the fingerprint of LiuWei DiHuang pills by near infrared spectroscopy].

The purpose of the present paper is to establish a method of the fingerprint of LiuWei DiHuang pills by the near infrared spectroscopy. First, the authors established the fingerprint of LiuWei DiHuang pills by high performance capillary electrophoresis (HPCE), and calculated its similarity. At the same time, the authors scanned its near infrared spectrogram. Then the authors established the mathematical model between the similarity of fingerprint and the near infrared spectrogram. Through the optimization of the model, the correlation(r), calibration standard deviation and the average relative error of the modeling set were 0.9046, 0.058 and 6.12%, respectively. It proved that the linearity between calculated and forecast of fingerprint was clear. The results showed that the method of the fingerprint of LiuWei DiHuang pills by the near infrared spectroscopy was feasible.

Effects of blood-activating and stasis-removing drugs combined with VEGF gene transfer on angiogenesis in ischemic necrosis of the femoral head.

OBJECTIVE: To observe the promoting effects of blood-activating and stasis-removing Chinese drugs combined with vascular endothelial growth factor (VEGF) gene transfer on angiogenesis in ischemic necrosis of the femoral head. METHODS: Forty Japanese giant-ear rabbits were randomly divided into a control group, a model group, a Chinese drug group, a gene group, and a combined group. After 8 weeks of treatment, the rate of VEGF positive cell expression in the synovium of the femoral head was measured using the immunohistochemical method, and the number of blood vessels in the femoral head was measured by digital subtraction angiography. RESULTS: The rate of VEGF positive cell expression in the model group was significantly lower than that in the Chinese drug group (P < 0.05) and very significantly lower than those in other groups (P < 0.01); but in the combined group it was significantly higher than in the Chinese drug group (P < 0.05). The differences in the number of blood vessels in area A between the model group and other groups were not statistically significant. However, in area B, the number of blood vessels significantly increased in the control group, the gene group and the combined group as compared with the model group (P < 0.05), and in the combined group the number of blood vessels was significantly more than in the gene group (P < 0.05); but in the Chinese drug group it was not significantly different than the model group (P > 0.05). CONCLUSION: Either the blood-activating and stasis-removing Chinese drugs or VEGF gene transfer can promote the angiogenesis and building of collateral circulation for femoral head ischemic necrosis, and the combined therapy with Chinese drugs or VEGF gene transfer may show a better therapeutic effect. The present study provides an experimental basis for clinical application of the combined therapy with the blood-activating and stasis-removing Chinese drugs and VEGF gene transfer.

[Study on the projection method based on principal component of tobacco near-infrared spectrum and application in redrying model].

The present paper briefly describes the application of near-infrared spectroscopy technology in tobacco. Two methods for solving projection vector based on the principal component of near-infrared spectrum are reported. They are named as projection of basing on principal component and Fisher criterion by principal component analysis method (PPF_PCA for short) and projection of basing on principal component and Fisher criterion by solving eigenvalue (PPF_Eig for short), and they are studied and compared in the application and evaluation of the redrying model. The result of the first-dimensional projection on 9 kinds of tobacco leaf grading samples shows that, the diversity of the first-dimensional projection values of inter-class and intra-class by the PPF_PCA method is both larger than that by the PPF_Eig method, and the mean absolute deviation of the mean projection values of inter-class by the PPF_PCA method is about 1.26 times that of the PPF_Eig method. At the same time, this result is interpreted by using the contribution rate of the first-dimensional projection values. That is, the contribution rate of first-dimensional projection values by the PPF_PCA method is 93%, while the contribution rate of first-dimensional projection values by the PPF_Eig method is 77%. The former is about 1.21 times the later. Therefore, the first-dimensional projection values by PPF PCA method include more information of diversity of both inter-class and intra-class. The similarity of samples inter-class and the diversity of samples intra-class can be evaluated more objectively from first-dimensional projection figure(on 9 kinds of tobacco leaf grading samples, 33 kinds of tobacco leaf grading samples and 6 redrying blending models), so it is more convenient to be used as a reference for the redrying model of tobacco, and it has a good application prospect in other formulation design of agricultural products (traditional Chinese medicine etc.).

[Application of wavelet packet entropy and Fisher classifier to the identification of medicinal rhubarbs with near-infrared spectrum].

The diffused-reflectance near-infrared (NIR) spectrum of medicinal rhubarbs was collected by Fourier transform spectroscopy instrument Principal components (PC) and wavelet packet entropy (WPE) were then calculated from the spectrum. Based on these two kinds of features, the models of identification of medicinal rhubarbs were developed using Fisher classifier. The results show that the error rates of cross-validation and prediction using WPE are all lower than those using PC. The model was built by WPE feature extraction method combined with Fisher classifier, the error rate of cross-validation is 6.52%, while that for prediction is 2.04%. The research result provides a method for identifying medicinal rhubarbs quickly.

[Influence of sample loading and test conditions on NIR veracity and study of analysis error source].

The influence of sample test conditions on the NIR veracity was studied with homemade grating diffuse NIR instrument using Yunnan flue-cured tobacco. Deducing analysis error was achieved by model self-emendation when a global NIR model was set up. Without regarding the influence of loading samples and test conditions, the test repetition error, re-loading error and samples tightness error, which were brought by instrument S/N, accounted for 50%, 30% and 20% of the total error, respectively. Depressing sample could reduce errors brought by sample tightness. Changes in test conditions could bring more analysis error, which was larger than the total of repetition error. These results theoretically explain the influence of sample test conditions on the NIR analysis veracity, which can provide basic theory data for farther improvement of homemade instrument and offer a new idea for resolving this problem.

[Influence of sample annum and distribution of chemical values on NIR veracity].

The influence of sample annum and the distribution of sample component on NIR veracity was studied with homemade grating diffuse NIR instrument using Yunnan flue-cured tobacco. Results showed that sample annum had an obvious influence on the total sugar and nicotine models, but had an unconspicuous influence on the total-nitrogen model. Models set up by samples, whose component content distribution was normal school, was better than those set up by even distribution. The conclusion in this study has a significant referenced value for the method and principle to select representative samples to modeling from a large amount of specimens.

[New technology for sample preprocessing in NIR analysis].

Sample's moisture content and granularity would have an important influence on NIR veracity. Preprocesssing methods have been used for NIR analysis to achieve better NIR veracity. With Yunnan flue-cured tobacco as study materials, the authors innovatively applied microwave fast drying technology and home pulverizer to tobacco preprocessing, which could finish the preprocessing in 5 min, faster than the traditional methods such as oven drying and sifting. Results showed that: microwave fast drying technology could improve NIR analysis veracity when samples' moisture content had a wide range; influence of samples' granularity on NIR veracity could be eliminated after 25 s home pulverizer shatter. This study can offer a new method for NIR analysis workers to simplify NIR preprocessing.

[Predicting the chemical composition of intact kernels in maize hybrids by near infrared reflectance spectroscopy].

Intact-kernel samples of normal maize inbred lines and hybrids were collected from field experiments of three locations. Calibration equations were developed by partial least square regression (PLS) of chemical values of near infrared reflectance spectroscopy (NIRS) data and tested through both cross and external validation. In addition, 40 progenies of F1 and F2 generation not included in calibration and validation sets were verified to further evaluate the reliability of three calibration equations. The authors found the coefficients of correlation (r) of 0.98, 0.93 and 0.97 between NIRS predicted and actual protein, starch and oil content in these materials, respectively. However, the greatest relative errors were 2.7% (protein), 2.46% (starch) and 7% (oil). Thus, the accuracy of prediction could be comparable to chemical methods. The feasibility of developing NIRS equations with samples of inbred lines to determine grain quality of hybrids was also examined. The analysis of principal components of spectrum of the inbred lines and hybrids supported a new theory that plant spectrum properties could be heritable.

[Study on application of Fourier transformation near-infrared spectroscopy analysis with support vector machine (SVM)].

Support Vector Machine (SVM) is a method for the research on identifying two types of problem. It is the latest branch in the statistics study theories, and the identification model has a strict mathematics foundation. In this paper, the basic principle and method of SVM are not only introduced, but also applied to chemometrics. One hundred and three rhubarb samples were used as experimental materials. The identification models were established with near-infrared spectroscopy and SVM training method with the intention of identifying whether the rhubarb samples are true or false. The thirty-three samples in training set were identified by the identifying models with the accurate rate of 100%, while seventy estimate samples had an accurate rate of 96.77%. The research result provided the method of identifying the traditional Chinese medicine rhubarb quickly. So, it shows the feasibility of establishing the models with near-infrared spectroscopy and SVM method to identify biological samples. This paper introduced the theme of SVM training method in order to beget the attention of the research members who deal with chemometrics.