For solving the problem that common similarity algorithms are insensitive to the relative differences of different chromatographic fingerprints data, this paper improved the algorithms of improved extent similarity), which named new improved extent similarity. It was confirmed that new improved extent similarity could distinguish) the samples with the same total difference and different distribution of relative differences. by applying) the similarity algorithm to simulated data and the chromatographic fingerprints of nine batches Rukuaixiao tablets. New improved extent similarity could reflect the relative differences of peak areas more sensitively when the relative differences of common peak areas was less than 100%. The result indicates that new improved extent similarity can reflect the large deviation from the established proportion of components, which can be applied to the control of the fluctuation of the established proportion of effective components and the process quality control of Traditional Chinese Medicine .

Near infrared chemical imaging (NIR-CI) is an emerging technology for rapidly analyzing the critical quality attribute of Chinese materia medica (CMM). It integrates NIR spectroscopy with chemical imaging. In this paper, it provided a systematic introduction to NIR-CI, such as the core part of instrument, the reliability, transformation, analysis and application of high-dimensional data acquisition. In addition, current studies of NIR-CI application in pharmaceutical field were analyzed. Finally, future opportunities and challenges of NIR -CI applications in the quality control of CMM preparation were prospected.

Objective:To establish a method for determination of total flavonoids,eriodictyol-7-methylether and isosakuranetin in Artemisia halodendron seeds.Method:The content of total flavonoids was determined by ultraviolet spectrophotometry at 506 nm,and compared with eriodictyol-7-methylether as a reference substance;the content of eriodictyol-7-methylether and isosakuranetin was determined by RP-HPLC,the analytical column was YMC-Pack ODS-A column(5?m,4.6 mm?250 mm),the mobile phase consisted of acetonitrile-0.2% phosphoric acid aqueous(35:65) at a ow rate of 1.0 ml?min-1,and the detection wavelength was 288 nm.Results:The linear range of total avonoids was 0.1228--0.7368mg(r=0.9991) and the average recovery(n=6) was 100.5%(RSD=1.41%);the linear ranges of eriodictyol-7-methylether and isosakuranetin were 3.016--30.16?g/ml(r=0.9999)and 3.304--33.04?g/ml(r=0.9999),respectively;the average recoveries(n=6) were 98.9%(RSD=1.20%)and 96.8%(RSD=1.38%),respectively.Conclusion:The methods of UV and HPLC are simple,accurate with good reproducibility and suitable for determination of total avonoids,eriodictyol-7-methylether and isosakuranetin.

To obtain active protein of pIL-18 expression in Lactococcus lactis, and to observe its biological activity, the total RNA was extracted as template from peripheral blood mononuclear cells. Porcine interleukin 18 (pIL-18) was amplified by RT-PCR. The resulting fragment was cloned into pAMJ399 L. lactis vector, and then transformed to L. lactis MG1363 cells by electroporation. Expression of pIL-18 protein was detected by SDS-PAGE and Western-blotting. Bioactivity of the product was tested by pig spleen lymphocyte proliferation test and cytopathogenic effect inhibition assay. The result of Western blotting and bioactivity test shows that the molecular weight of pIL-18 protein was 19 kDa. The react line was observed in both supernatant and precipitated of the recombinant bacteria pAMJ399-pIL18/MG1363. The expressed pIL-18 can promote the proliferation of pig spleen lymphocyte, and significantly inhibit virus multiplication. As conclusion, porcine interleukin-18 was successfully expressed in L. lactis, and the product was biologically active.
Animals ; Blotting, Western ; Electrophoresis, Polyacrylamide Gel ; Electroporation ; Genetic Vectors ; Interleukin-18 ; biosynthesis ; Lactococcus lactis ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Swine

The coding sequence for the mature peptide of porcine lactoferrin (Plf) was synthesized according to the codon usage of lactobacillus, to establish optimized porcine lactoferrin Lactobacillus expression system. The gene was ligated into the Xho I/BamH I site of Lactobacillus expression vector pPG612.1 and the recombinant plasmid pPG612.1-plf was transformed individually into Lactobacillus casei ATCC393, Lactobacillus pentosus KLDS1.0413, Lactobacillus plantarum KLDS1.0344 or Lactobacillus paracasei KLDS1.0652 by electroporation. After induction with xylose, expression of the recombinant proteins was detected by Western blotting and confocal laser scanning microscopy. Secretion of recombinant Plf proteins from four recombinant species was determined quantitatively by ELISA. The antibacterial activities of recombinant proteins were measured by agar diffusion method. The result shows that Plf was correctly expressed in four species of recombinant lactobacillus, with molecular weight of about 73 kDa. The expression levels in recombinant Lactobacillus casei, Lactobacillus pentosus, Lactobacillus plantarum, Lactobacillus paracasei were 9.6 μg/mL, 10.8 μg/mL, 12.5 μg/mL and 9.9 μg/mL, respectively. Antimicrobial activity experiment shows that the recombinant proteins were active against E. coli, Staphylococcus aureus, Salmonella typhimurium, Listeria, Pasteurella. The recombinant Plf expressed by recombinant Lactobacillus plantarum showed the best antibacterial activity among all recombinant lactobacillus species. These data represent a basis for the development and application of porcine lactoferrin from recombinant lactobacillus.
Animals ; Anti-Bacterial Agents ; biosynthesis ; Lactobacillus ; metabolism ; Lactoferrin ; biosynthesis ; Recombinant Proteins ; biosynthesis ; Swine

A new non-destructive and rapid method was developed to discriminate the coating process of Rukuaixiao tablets mainly based the support vector machine (SVM) with the near-infrared spectroscopy (NIRs). After the samples that differ in the sugar-coat were acquired, the sample set partitioning based on joint x-y distance (SPXY) method was used to select the training sets and internal principal component analysis (internal PCA) was to select the optimal wavelength. The discrimination model was developed based on support vector machine (SVM), and varieties of pre-processing methods were compared. The results showed that the accuracy of the prediction set was 98.81%. It is concluded that the accuracy of the method is high to use for the quality evaluation of tablet's coating process.
Algorithms ; Clinical Laboratory Techniques ; standards ; Clinical Protocols ; standards ; Drugs, Chinese Herbal ; analysis ; Medicine, Chinese Traditional ; standards ; Medicine, East Asian Traditional ; methods ; Principal Component Analysis ; Spectrophotometry, Infrared ; methods ; Spectroscopy, Near-Infrared ; methods ; Tablets ; analysis

This study was aimed to optimize the near infrared (NIR) variable selection method based on multivariate detection limit (MDL). Using Qing-Kai-Ling (QKL) injection as object, three variable selection methods (interval par-tial least-squares, iPLS; backward interval partial least squares, BiPLS; moving window interval partial least squares, mwPLS) were used to establish the PLS models of baicalin in QKL injection, respectively. The prediction ability of different variable selection method was compared. MDL of all models were calculated in contrast to the MDL value of full spectra PLS model, to select optimal variable selection method. The results showed that different variable selec-tion methods had different prediction ability. Among them, iPLS had the best performance which determination coef-ficient of prediction (Rpre2) and the root mean square errors of prediction (SEP) were 0.996 5 and 602.3 μg·mL-1, re-spectively. All MDLs of different variable selection methods were reduced compared with the full spectra PLS model. The value of iPLS was the lowest comes to be 1.19 μg·mL-1. The results above indicated that the best variable se-lection method for baicalin in QKL injection was iPLS. MDL theory took the error of calibration and validation set and the leverage of external sample into account, which can comprehensively evaluate model detection performance compared to the classic chemical indicator parameters. This method was particularly suitable for the variable selec-tion method optimization of NIR quantitative model of low concentration sample such as Chinese herbal medicine.

This article was aimed to study the different clinical characteristics using drug pair of Cassia twig and white peony root with the contents ratio of 1:1 and 1:2. Based on the different clinical treatment of drug pair of Cas-sia twig and white peony root, different compositional ingredients in ratio of 1:1 and 1:2 were illuminated by HPLC/MS method. The drug pair of Cassia twig and white peony roots in ratio of 1:1 and 1:2 and single herbs were ex-tracted for HPLC/MS analysis. A protocol was followed, including acetonitrile - 0.1% acetic acid with gradient elution, positive mode, 350℃ capillary temperature and 300℃ vaporization temperature. The results showed that Procyanidol B2 and 2-Hydroxy cinnamal dehyde can be extracted from single Cassia twig, but 2-Hydroxy cinna-mal dehyde cannot be detected in drug pair. It showed the contents of Procyanidol B2 in 1:1 ratio was more than 1:2 ratio. Simultaneously, Palbinone, paeoniflorin sulfonate, 1,2,3,6-Tetra-O-galloyl-β-D-glucose, Paeoniflorin, Pae-oniflorin isomers, Benzoylpaeo-niflorin, and Benzoyl Paeoniflorin isomers can also be dissolved in white peony root. In addition, the contents of 1,2,3,6-Tetra-O-galloyl-β-D-glucose, Paeoniflorin, Benzoylpaeo-niflorin, and Benzoyl Paeoniflorin isomers in 1:1 were more than 1:2. The contents of Palbinone, paeoniflorin sulfonate and Paeoniflorin isomers in 1:2 were more than 1:1. It was concluded that Procyanidol B2, 1,2,3,6-Tetra-O-galloyl-β-D-glucose, Paeoniflorin, Benzoylpaeo-niflorin and Benzoyl Paeoniflorin isomers in 1:1 were more than 1:2. The contents of Pal-binone, Paeoniflorin sulfonate and Paeoniflorin isomers in 1:2 were more than 1:1. It provided a scientific basis for traditional Chinese medicine treatment using rational drug pair.

ToprovidethemethodologyforrapidqualityevaluationofLonicerajaponica,wehaveestablished the stable quantitative model of near infrared spectroscopy ( NIR) . The performance of Bagging partial least squares (Bagging-PLS) model and Boosting partial least squares (Boosting-PLS) model was compared with that partial least squares ( PLS ) model based on the NIR data of ethanol precipitation process of Lonicera japonica. On this basis, the performance of these two models after variables selection was also studied by the methods of siPLS ( synergy interval partial least squares ) and CARS ( competitive adaptive reweighted sampling) . The experimental results showed that the prediction performance of Bagging-PLS and Boosting-PLS models was superior to PLS model with the latent factor of 10 . The band of 820-1029 . 5 nm and 1030-1239. 5 nm for the first batch was selected by the method of siPLS. In addition, the band of 820-1029. 5 nm and 1030-1239. 5 nm was selected for the second batch sample in the same method. Furthermore, the method of CARS was taken to select variables for the two batches samples with 5-fold cross-validation and 10-fold cross-validation. And the lowest RMSECV( root mean square error of cross-validation) values were used to take subset. Compared to the model performance without the method of CARS, the RMSEP value of the Bagging-PLS model and Boosting-PLS model for the concentration of chlorogenic acid reduced by 0 . 02-0 . 04 g/L and rp(correlation coefficient of prediction)value increased by 4%-5%. Generally, Bagging-PLS and Boosting-PLS could be regarded as rapid prediction methodsfor NIR quantitative models of ethanol precipitation process of Lonicera japonica.

Laser-induced breakdown spectroscopy (LIBS) was applied to perform qualitative research on Tibetan medicine “GTso Thal” in order to establish a rapid element analysis method . The Nd: YAG laser with the funda-mental frequency at 1064 nm was used. A high-power laser beam was focused on the surface of the sample. The spectrometer and CCD was used to detect its spectrum signals. Based on the National Institute of Standards and Technology (NIST) database, emission spectrum characteristics were differentiated. The LIBS spectra of “GTso Thal”showed multi-elements including Hg, Ca, Na, As, Fe, Mg, Al, K, Li, Pb, Ag, Au and S. Among them, Hg, Pb, Ag and Au were heavy mental elements. The results demonstrated that LIBS was a viable technique for analysis of Ti-betan medicine “GTso Thal”. LIBS provided reliable elemental analysis on Tibetan medicine “GTso Thal”. The de-tection was real-time, rapid and in situ. It had prospects in the elemental analysis of ethnic medicine study. LIBS had broad application prospects.