[Study on Panax notoginseng and its processed products by FTIR spectroscopy].

The chemical differences of panax notoginseng before and after processing were analyzed by Fourier transform Infrared spectroscopy (FTIR) combined with two-dimensionalcorrelation spectroscopy (2D-IR). Compared with conventional IR spectra of the samples, the FTIR spectra of panax notoginseng and its processed products were similar in the range of 1 200-400 cm(-1). The difference was that prepared panax notoginseng had strong and characteristic peaks at 2 925, 2 855, 1 746, 1 460, 1 376 and 1 158 cm(-1), which all arose from the characteristic vibration of peanut oil. This was because there was some peanut oil left in the panax notoginseng, when panax notoginseng after processing. Obvious differences were observed between 2D-IR spectra of them, in the range of 1 400-1 700 cm(-1), there was only one auto peaks at 1 650 cm(-1) in the spectra of panax notoginseng, but there were auto peaks at 1 469 and 1 640 cm(-1) in the spectra of prepared panax notoginseng. In the range of 1 400-1 700 cm(-1), the 2D-IR spectra of panax notoginseng and its processed product present characterstic peaks at 1 139 (1 137), 1 194 (1 196), 1 121 (1 221)cm(-1) respectively, but the relative intensities of auto peaks were changed. For example, auto peak around 1 139 cm(-1) was enhanced, but auto peak around 1 194 cm(-1) was weakened. The results of 2D-IR correlation spectroscopy indicated the decomposition of flavonoids, saccharides and saponins. This method can track dynamically the processing procedure of panax notoginseng and reveal the main tansformations, so it can explain the pharmacology of panax notoginseng and its processed product by FTIR and 2D-IR.

[Study on infrared spectrum change of Ganoderma lucidum and its extracts].

From the determination of the infrared spectra of four substances (original ganoderma lucidum and ganoderma lucidum water extract, 95% ethanol extract and petroleum ether extract), it was found that the infrared spectrum can carry systematic chemical information and basically reflects the distribution of each component of the analyte. Ganoderma lucidum and its extracts can be distinguished according to the absorption peak area ratio of 3 416-3 279, 1 541 and 723 cm(-1) to 2 935-2 852 cm(-1). A method of calculating the information entropy of the sample set with Euclidean distance was proposed, the relationship between the information entropy and the amount of chemical information carried by the sample set was discussed, and the authors come to a conclusion that sample set of original ganoderma lucidum carry the most abundant chemical information. The infrared spectrum set of original ganoderma lucidum has better clustering effect on ganoderma atrum, Cyan ganoderma, ganoderma multiplicatum and ganoderma lucidum when making hierarchical cluster analysis of 4 sample set. The results show that infrared spectrum carries the chemical information of the material structure and closely relates to the chemical composition of the system. The higher the value of information entropy, the much richer the chemical information and the more the benefit for pattern recognition. This study has a guidance function to the construction of the sample set in pattern recognition.

[Infrared spectroscopic analysis of Guilin watermelon frost products].

The objective of the present study is to analyze different products of Guilin watermelon frost by Fourier transform infrared spectroscopy (FTIR), second derivative infrared spectroscopy and two-dimensional correlation spectroscopy (2D-IR) under thermal perturbation. The structural information of the samples indicates that samples from the same factory but of different brands had some dissimilarities in the IR spectra, and the type and content of accessories of them were different compared with conventional IR spectra of samples, peaks at 638 and 616 cm(-1) all arise from anhydrous sodium sulfate in watermelon frost spray and watermelon frost capsule; the characteristic absorption peaks of the sucrose, dextrin or other accessories can be seen clearly in the spectra of watermelon frost throat-clearing buccal tablets, watermelon frost throat tablets and watermelon frost lozenge. And the IR spectra of watermelon frost lozenge is very similar to the IR spectra of sucrose, so it can be easily proved that the content of sucrose in watermelon frost lozenge is high. In the 2D-IR correlation spectra, the samples presented the differences in the position, number and relative intensity of autopeaks and correlation peak clusters. Consequently, the macroscopical fingerprint characters of FTIR, second derivative infrared spectra and 2D-IR spectra can not only provide the information about main chemical constituents in medical materials, but also analyze and identify the type and content of accessories in Guilin watermelon frost. In conclusion, the multi-steps IR macro-fingerprint method is rapid, effective, visual and accurate for pharmaceutical research.

[Similar spectrum of infrared spectrum and its application in identification of Chinese herbs].

In the present paper, the similar spectra of 18 samples, which include Astragalus, red-blue Astragalus and Codonopsis, were obtained in the range of 1600-700 cm(-1). The result showed that all kinds of herbs have their own characteristic similar spectra, and 18 samples can be identified according to the characteristic similar spectra. Furthermore, three correlation coefficients of 93 ganoderma samples were calculated which is in the range of 1560-1502, 1460-1421 and 1319-1260 cm(-1) according to the information of similar spectrum of infrared spectrum of ganoderma. Without priori knowledge of the classification of these samples, the K-means cluster analysis can successfully divide them into four classes, i.e., Ganoderma lucidum and Ganoderma sinensis, Ganoderma atrum, Ganoderma aoshiba, Ganoderma multiplicatum. This result is consistent with the result of morphological classification.

[Analysis and identification of Huangqi and its counterfeit ciguogancao by two dimensional corrlation infrared spectroscopy].

Standard Huangqi (Astragalus membranaceus (Fisch) Bge. Var. monghlicus (Bge.) Hsiao) and its counterfeit Ciguogancao (Glychrrhiza pallidiflora Maxim.) can be discriminated and identified by using multi-steps infrared maro--fingerprint method. In the 1D-IR spectra, the peak intensity at 1 737 cm(-1) for Ciguogancao, which is the stretching vibration peak of C==O, is much stronger than that of Huangqi. It's proved that the organic ester compounds in Ciguogancao are much more than Huangqi. In the secondary derivative spectra, it's easy to find the fingerprint characteristic peaks of CaC2O4 in the infrared spectra of Ciguogancao, but not in that of Huangqi. Besides, both secondary derivative spectra also have main characteristic peaks, which are the skeletal stretching of aromatic, round 1463, 1511 and 1596 cm(-1), but Ciguogancao aslo has one shoulder peak at 1 453 cm(-1). In the 2D-IR spectra, both have three auto-peaks at 1070, 1095 and 1140 cm(-1), which are the autopeaks of glucoside, but the strongest auto-peak of Huangqi is at 1140 cm(-1) and that of Ciguogancao's is at 1 090 cm(-1). The spectra testified that the organic ester compounds, aromatic compounds and glucoside compounds in Huangqi and its counterfeit Ciguogancao were different. The method not only can identify standard Huangqi and its counterfeit Ciguogancao rapidly, but also provides useful information about the differences in organic ester compounds, aromatic compounds and glucoside between Huangqi and its counterfeit Ciguogancao. It's proved that multi-steps infrared maro-fingerprint method can be used to analyze and distinguish Huangqi and its counterfeit Ciguogancao.

[Application of comparison method with correlation coefficient array in identification of Chinese herbs].

The infrared spectrum of Chinese herbs possesses the character similar to fingerprint. The purpose to identify Chinese herbs can be achieved by way of comparing these spectra. The software of comparing correlation coefficient array of FTIR spectra was designed in Visual Basic 6.0. It established an infrared spectrum database including more than 300 kinds of Chinese herbs. The FTIR spectra of unknown herb can be compared with those of Chinese herbs in the database, and this method can identify the suitable herbs according to the similar herbs in the database. The study shows that within the spectral database of herbs, the factors of spectral interference, such as H2O, CO2, etc., can be avoided through assigning threshold values of correlation coefficients and comparing spectra with characteristic peak bands, and the comparison result is credible and correct. This method was expected to be a common method in identifying Chinese herbs.

[Recognition of three classes of skullcaps by FTIR spectroscopy combined with artificial neural networks].

In order to recognition of three classes of skullcaps (cultivated, wild Scutellaria baicalensis Georgi and Scutellaria viscidula Bge) three kinds of models of artificial neural networks (ANN), nonlinear-linear, linear-linear and nonlinear-nonlinear model, were used combined with their infrared spectra. Skullcaps samples were collected by Fourier Transform Infrared (FTIR) spectra. 42 samples were gathered as a train set, and 34 samples as a test set, then their supervision trains were performed using three models each. When the summation of error square of train target was selected as 0.01, the correct rate for recognition of three classes of skullcaps using each ANN was 100% for the train set, but was different for the test set, which depended on the number of node in hidden layer, S1. It was found that with the increase of S1, the correct rate would decrease oppositely. This may be caused by the high degree of the non-linearity of the networks, so that the models of networks were not fit for the train of this kind of sample set. When using linear-linear model of ANN varied with S1 in some extent, the correct rate was generally about 85%. Recognizability obtained using nonlinear-linear model of ANN was the best. Its correct rate of recognition was > 97% when S1 = 3, and so this method can be used to recognize three of skullcaps simply, rapidly, and accurately.

[Quick identification of scullcaps in different geographical origins using clustering analysis method and infrared fingerprint spectra].

Based on the infrared fingerprint spectra, scullcap samples from 15 geographical origins were clustered into 6 classes with principal component analysis. The classification was related to their geographical origins and weather. In the same class, chemical components of scullcaps are similar to each other, which can be considered as the criterion of evaluating the quality of scullcaps. 43 geographical origins were predicted with radial basis function, which was demonstrated to be a powerful ANN method in discrimination. The classes of scullcaps can be distinguished by the method. The method can be used to class and identify geographical origins of scullcaps. It is one of the methods in the quality control of traditional Chinese medicines.

[Expression of WT1 gene and its isomer ratio changes during phorbol ester induced differentiation of K562 cell line].

OBJECTIVE: To explore the changes in expression of WT1 gene and ration of its isomers during phorbol ester (TPA) induced differentiation of leukemia cell line K562 by fluorescence quantitative RT-PCR and analysis the relationship between different isomers and hematogenic cell differentiation. METHODS: The degree of cellular maturation were verified by NBT reduction test and immunophenotyping. Expression of WT1 gene was determined by fluorescence quantitative RT-PCR during differentiation of K562 cell line. The relative ratio of the four splicing variants WT1 ( + / + ), WT1 ( + / - ), WT1 ( - / + ), WT1 ( - / - ) were calculated. RESULTS: During the differentiation of K562 cell, the NBT reduction rate and the CD9 positive rate both increased significantly (P < 0. 05). The expression of WT1 gene decreased immediately to (1.67 +/- 0.45) x 10(-3) from (4.67 +/- 1.11) x 10(-3), and then increased again to (4.64 +/- 1.53) x 10(-3) at 96 hours. The ratio of WT1 ( + / + ) was decreased gradually, from 0 hour (39.65 +/- 19.46)% to 96 hour (15.25 +/- 7.27)%. While the ratio of WT1( - / - ) was increased, from 0 hour (15.38 +/- 11.34)%, to 96 hour (37.60 +/- 11.90)%. The other two isomers ratios did not change significantly. CONCLUSION: During the TPA induced differentiation of K562 cell, there are two high expression levels of WT1 gene. Before differentiation, the majority is WT1 ( + / + ), and after differentiation, is WT1 ( - / - ). It indicates that WT1 gene may activate or inhibit cell differentiation by regulating the ratio of its four splicing variants.

[Fluorescence quantitative PCR detection of WT1 gene expression in peripheral blood of patients with acute leukemias and its clinical implications].

To elucidate the expression of WT1 in all types of leukemias and its implications for monitoring minimal residual disease in patients with acute leukemia, the peripheral blood from 55 leukemia patients and 10 normal voluteer was detected by using FQ-RT-PCR. Follow-up monitoring of WT1 expression of peripheral blood was performed for 20 patients with acute leukemia. The results showed that the expression of WT1 gene in all types of leukemias was significantly higher than that in normal control (P < 0.001). For ANLL and ALL patients, the survival time in the group of WT1 6.8 x 10(-3), (P = 0.027). Follow-up detection of the expression of WT1 in peripheral blood samples from 20 acute leukemia patients, 7 cases relapsed after complete remission has been done. In 5 of 7 relapsed patients, the expression of WT1 had obviously increased about 2 - 3 months before clinical relapse became apparent. It is concluded that the established FQ-RT-PCR method is accurate and specific. The expression of WT1 gene is relatively high in all types of leukemias compared with normal peripheral blood cells, the higher WT1 expression may associate with poor prognosis in acute leukemia, and the dynamics of WT1 level correlate with the disease status. The quantitative assessment of WT1 expression in peripheral blood samples by FQ-RT-PCR may be a useful tool for monitoring minimal residual disease.