[Screening of Shuanghuanglian Injection allergenic ingredients based on immune fingerprint].

In this paper,immune fingerprint was used to screen the allergenic components of Shuanghuanglian Injection(SHLI) by enzyme-linked immuno sorbent assay(ELISA) combined with HPLC/MS method. ELISA-embedded anti-IgE antibody could successfully adsorb allergens in SHLI and its plasma samples containing drugs through different routes of administration,suggesting that SHLI can induce type I hypersensitivity in rats. HPLC fingerprints and MS map of SHLI and drug-containing plasma samples from different routes of administration before and after anti-IgE antibody adsorption were established. According to the similarity evaluation of HPLC fingerprints and analysis results MS map,the sensitization of traditional Chinese medicine injections can be changed by different administration methods. There were 22 kinds of components that can be adsorbed by specific anti-Ig E antibodies in Shuanghuanglian Injection and its drug-containing plasma,most of them were acids and nitrogen compounds. Based on supramolecular theory,it was inferred that these compounds came from SHLI or body,and may form supramolecular hapten,which results in immunotoxicity and allergic reaction when being used as injection instead of oral liquid. Immune fingerprint is not only used to screen out single component allergen,but also more comprehensive,sensitive and easy to operate. It can provide reference for the future research methods of allergic reaction of traditional Chinese medicine injections.

[Research status of common processing technology of traditional Chinese medicine "maintaining medicinal properties after carbonisatus" and supramolecular "imprinting template" characterization technology].

Based on the characteristics of biological supramolecules and the law of " imprinting template",the research status and common problems in " maintaining medicinal properties after carbonisatus" in traditional Chinese medicine( TCM) were analyzed,and the further countermeasures were put forward. According to the historical evolution of " maintaining medicinal properties after carbonisatus" in TCM processing,the origin of its common problems was clarified by using the theory of biosupramolecular chemistry. TCM is a megacomplex biological supramolecular system,so TCM processing is just the processing of megacomplex biological supramolecular system,and its essence is a TCM pharmaceutical technology with chemical changes in host and guest of biological supramolecular system with or without adjuvant material under high temperature and humidity. In this study on pharmaceutical technology,host molecule was destructed in the process of carbonizing,but guest molecule was retained. The changing law of the host and guest molecule was controlled by the " imprinting template",which was reflected in the degree of change in the drug properties and efficacy of the decoction pieces. Supramolecular chemistry ran through the whole process,and the " imprinting template" of charcoal medicine was characterized by the supramolecular topological structure characteristics and imprinting behavior. After being combined with the quantitative mathematical model of heating degree in processing,it can realize the accurate processing of " maintaining medicinal properties after carbonisatus" from the source,quantitatively control the quality of carbonic herbs,and formulate stable and controllable quality standards.

[Q-marker of Chinese medicine and metabolic regularity of co-network compatibility and rainbow potential].

The effect of Chinese medicine (CM) compound prescription is the combined action results of single herbs based on the basic theory of Chinese medicine, and its function embodies the characteristics of multi components, multi targets and comprehensive effects. It is difficult to study the therapeutic material, establish quality standards or determine Q-marker, so we can't strictly monitor the quality of the whole process of CM. The identification of Q-marker has a profound influence on the whole process of the pharmaceutical engineering of CM. The scattered effect of CM multi-components is regarded as the integral action of the parent nucleus group by the metabolic rule of co-network compatibility and rainbow potential (CCRP). The rule can be used to communicate the individual components and macro components, to reveal the metabolism of CM in organism body and basic law of information exchange, thus revealing the action law of CM on human body. Through the systematic analysis of the Q-marker's guidance to the development of CM and its relationship with the metabolic rule of CCRP, we try to provide some ideas for the identification of Q-marker.

[Solubilization characteristics of licorice based on supramolecular "imprinted template" theory].

To investigate the effect of the ingredient group of traditional Chinese medicine (TCM) with different "imprinted template" on the supramolecular solubilization ability of licorice, in order to lay a theoretical foundation for explaining the solubilization phenomenon of the components of TCM. Based on the independent supramolecular "imprinted template" rules, molecular connectivity index (MCI) and the correlation of n-octanol-water partition coefficient (logP) were used to indicate hydrophilic lipophilic capacity of TCM, and the extractum rate was used to indicate the solubilization effect of licoriceon single TCM herbs or compounds. The solubilization ability of licorice was evaluated based on MCI, logP and the extractum rate. According to the results, the correlation coefficient between MCI and logP for single herbs was 0.942, and that for single herb adding licorice was 0.916. The extractum rate of most herbs was increased after adding licorice. The correlation coefficient among the extractum rate as well as MCI and logP change values before and after adding licorice were respectively 0.837, 0.405. The correlation coefficient between MCI and logP for eight compounds was 0.937. Meanwhile, licorice had a solubilization effect on the remaining 7 compound except for Huangqi decoction. Therefore, licorice shows the solubilization effect through the independent supramolecular "imprinted template", so as to improve the hydrophilic lipophilic ability. There was a high positive correlation between the MCI and logP in ingredients for TCM, which could be used as important parameters to indicate the "imprinted template" feature for components of TCM. The study on the solubilizing effect of TCM with the supramolecular "imprinted template" theory was feasible, and will lay a foundation for the reform of single-herb dosage form.

[Investigation of supramolecular chemical mechanism of traditional Chinese medicine processing].

To analyze the research status of traditional Chinese medicine (TCM) processing and key scientific issues to be resolved by using characteristic of TCM biological supramolecules and reaction theories of 'Qi chromatography' to human being, and then put forward the solution countermeasures. Based on the previous explications of theories of the Chinese medicine with supramolecular chemistry as 'Qi chromatograpy', biological supramolecular chemistry theory was used to explain the root of the problem in TCM processing and form the countermeasures for supramolecular chemistry research in the processing of TCM decoction pieces. TCM is a huge complex biological supramolecular body, so the processing of TCM is in view of the processing of huge complex biological supramolecular body. In nature, it is a TCM pharmaceutical technology with chemical changes of the subject and object of biological supramolecular body with or without auxiliary materials, under the condition of high temperature and high humidity. Supramolecular chemistry was throughout the processing of TCM, so the use of supramolecular technology was appropriate in research. TCM decoction pieces were the product of subject and object molecular changes after supramolecular chemistry reaction, with changes in physical and chemical properties of the subject and object molecules, such as bound water overflow, chemical bond rupture, dehydration, carbonization, and reaction with auxiliary materials. Changes in drug property and drug efficacy after processing depended on the degree of subject and object "imprinting template" change, which can be measured by 'Qi chromatography' function of supermolecular imprinting template in human body. In this paper, the research ideas, methods, variety attribution, processing principle, quality standard and clinical application were analyzed to find out the sticking point of the problems, and form the countermeasures with supramolecular chemistry as the core. The processing of TCM is a chemical technology of biological supramolecular body based on the function of supermolecular 'imprinting template' (medical element) of human meridian-viscera, and its reaction products (decoction pieces) showed determinacy when used alone and compatibility when used together, which may constitute a compatible drug delivery system of TCM in clinical application.

[Investigation on supramolecular mechanism of multi-component overall control in Chinese medicine controlled sustained-release preparation].

Controlled sustained-release preparation (CSRP) is a definitely important source of innovative medicines, however, its further researches and development are limited due to the complexity of Chinese medicine (CM) compositions, and a series of unclear key problems like the apparent properties from physical and chemical aspects. Combined with the CM characteristics of the multiple components and synergistic effect, a series of bottleneck problems in the preparation of CSRP were analyzed. Then the mechanism and influences of supermolecular chemical theory in the CSRP about the multiple-components overall-control were explored under the guideline of "overall control, synchronous dissolution" in CSRP, based on the previous explications about supramolecular chemistry theories and with the application of single component precision controlling technologies. The supermolecular mechanism of multi-component overall control and synchronous release of the CSRP was investigated, providing an useful academic reference for the research and development of the CSRP and laying the theoretical and technical foundation for the manufacturing process of CSRP.

[Impact of ″imprinting templates″ characteristics of traditional Chinese medicine injection supramolecules on (anaphylactoid) hypersensibility].

The (anaphylactoid) hypersensibility mechanism of ″imprinting templates″ characteristics of traditional Chinese medicine (TCM) injection supramolecules was clarified to lay the foundation to build safety evaluation method. Based on the previous publication on special impact of Chinese medicine theories on supramolcular chemistry, combined with the natural origination of (anaphylactoid) hypersensitized special rules as well as the sensitization phenomenon of cordate houttuynia injection, the impact of the structure characteristics of ″imprinting templates″ in TCM injection supramolecules on its (anaphylactoid) hypersensibility was clarified. In Chinese medicine injections, the supramolecular structures can independently be generated, showing overall apparent (anaphylactoid) hypersensibility nature, and their structure characteristics were dependent on the strength. In addition, (anaphylactoid) hypersensitive critical supramolecular structure was present. When it was administrated by ″injection″, it's structure was not easy to be destroyed, often showing apparent immunogenicity, whereas if it was administrated by ″oral″, the structure would be destroyed by the gastrointestinal tract, showing weaker or no apparent immunogenicity. Therefore, there are differences in (anaphylactoid) hypersensibility between ″injection″ and ″oral″ administration of TCM. TCM injections would produce the supramolecules between ″molecular society″ by independent reaction of supramolecular ″imprinting template″ (chemical determinants), showing apparent immune process of recognition, copying, and storage. Single molecule is a special example for this. The screening of anaphylactoid (sensitinogen) includes the single ingredients and their forming supramolecules for TCM injection. This is the unique feature for safety evaluation of Chinese medicine injection.

[Dispute for Japanese(wild) honeysuckle flower based on supermolecular imprinting templates].

More and more disputes have happened to confront us continuously since the separation of Japanese(wild) honeysuckle flower in Chinese Pharmacopoeia in 2005. The state pharmacopeia committee decided to separate Japanese(wild) honeysuckle flower into two species for japanese(wild) honeysuckle flower, but didn't define their the convincing reasons still did not provide to us as a result that two medicines are not described the differences in natural properties, efficiency and indication, usage and dosage, as well as not given a resolving methodand specific solution. It was known for us that in the history of traditional Chinese medicine（TCM）, the phenomenenphenomena of the "one drug from multi-species" and "one species for multi-drug" are very ordinary thingswere ubiquitous. Whether separation of the drug species are separated shall be decided to by clinical efficiency. Through Chinese pharmacopoeia (2015 edition) issue of Chinese Pharmacopoeia the 2015 issue of the Chinese Pharmacopoeia, we still cannot find a scientific solution for the dispute of for Japanese (wild) honeysuckle flower, perhaps because of insufficient reorganization of TCM clinical medication regularities, such as "treatment of different diseases with same drug", " treatment of the same disease with different drugs" and "treatment of the same syndrome with multi-prescriptions", and "one prescription treating multiple syndromes"; lack of in-depth analysis for multi-component TCM compounds and autonomisation of "supermolecular template" for organs and meridians; less attention to the advance of efficacy and safety evaluation technologies for multi-component TCM compounds; impacts from the medication mode of "one ingredient-one composition-one effect"; as well as insufficient research methods for bioequivalent evaluation in preclinic and clinic studies . The dispute for species combination or separation for Japanese(wild) honeysuckle flower was apparently caused by regional economy, drug biological equivalent of efficacy and safety, but arising from clinical principles for systematical syndrome treatment with TCM, or concepts in the treatment of diseases with TCM or western medicines. This paper focused on current studies on Japanese(wild) honeysuckle flower in the combination with TCM clinical medication regularities, such as "treatment of different diseases with same drug", "treatment of the same disease with different drugs" and "treatment of the same syndrome with multi-prescriptions", and "one prescription treating multiple syndromes", expounded the specific pharmacological regularity of "heterogeneous equivalence" of Japanese (wild) honeysuckle flower, and put forward methods for studying bioequivalence of Japanese(wild) honeysuckle flower, in order to solve the combination and separation of Japanese(wild) honeysuckle flower and lay a foundation for promoting the development of Chinese herbal medicine industry.

[Exploration of research approaches of Chinese medicine's pharmacology based on "imprinting templates" (medical element) of supramolecules].

The paper, based on the previous publication as special impact of Chinese medicine theories on supramolcular chemistry, aims to analyze the natural origination for the Chinese medicine and to explain the special impact of "Qi chromatography" reaction on "imprinting templates" in supramolcular host of human being with Chinese medicine, in order to reveal the CM's properties of "medical element" with "imprinting templates" autonomisation generally took place in natural supramolecules, and also to discover that the CM's pharmacology are satisfied with its own approaches different form western pharmacology. It was decided, for CM's pharmacology guided by CM's theories, to "Qi chromatography" relations between the CM's ingredient groups and the meridian zang-fu viscera. The supramolcular chemistry played an all-through role in procession of making macro-regularities and special presentation on behavior of "Qi chromatography" impulse owning to the matching action of all kinds of ingredients on the meridian zang-fu viscera with similar "imprinting templates". The CM's pharmacology were guided by CM's theories, owing to its interpretation of supramolecular chemistry. The pharmacology was achieved to construct up completely on base of classical chemical single molecular bonds whereas the CM's pharmacology be configured to big building by way of "imprinting templates" as multi-weak bonds among "supramolecular society". CM's pharmacology was supramolcular pharmacology dealt with "molecular society" on the base of western pharmacology, and employed to double research approaches both math-physical quantitative representation on macroscope and qualitative analyses in microscope.

[Study on meridian tropism of medicinal property theory for Chines medicines by supramolecular chemistry (I)].

In this paper, based on the special influence of supramolecular chemistry on the basic theory of Chinese medicines ( CM) , the authors further analyzed the history of meridian tropism and natural origins of CM organisms and explained CM ingredients and the universal regularity of the automatic action of the supramolecular "imprinting templates" hole channel structure. After entering human bodies, CMs, as the aggregation of supramolecular "imprinting templates" , automatically seek supramolecular subjects that are matched with their "imprinting templates" in human meridians and organs for the purpose of self-recognition, self-organization, self-assembly and self-replication, so as to generate specific efficacy in meridians and organs, which is reflected as the meridian tropism phenomena at macro level. This regularity can be studied by in vitro and in vivo experimental studies. In vitro methods are mostly supra molecular structure analysis and kinetic and thermodynamic parameter calculation; Whereas in vivo methods are dominated by the analysis on object component distribution, chromatopharmacodynamic parameters and network chromatopharmacodynamic parameters; Particularly, the acupoint-medicine method can simplify to study the supramolecular subject-object relations. Consequently, CM's'meridian tropism reveals the universal regularity for interactions of macromolecular and micromolecular "imprinting templates" of subjects and objects in natural organisms. As the first barrier for the material base of the CM theory and breakthrough in the modernization of the basic CM theory, meridian tropism plays an important role in studies on basic theories of the basic CM theory.

[Special impact of supramolecular chemistry on Chinese medicine theories].

The paper aimed to elucidate the specific impact of supramolecular chemistry on the Chinese medicine theories (CMT) in their modernization, after had summarized up the research status of supramolecular chemistry and analyzed the possible supramolecular forms of Chinese medicine (CM), as well as considered the problems in modernization of CM theories. On comparison of the classical chemistry that delt with chemical bonds among atoms, the supramolecular chemistry was rather concerned with varietes of weak noncovalent bonds intermolecules, and reflected the macro-apparent chemical properties of each molecules, and was the most appropriate chemical theories to explain the CMT and microcosmic materials. The molecules in the human body and Chinese material medica (CMM) formed supramolecules by way of self-assembly, self-organization, self-recognition and self-replication, with themselves or with complexation, composition, chelation, inclusion, neutralization etc. Meridian and Zang-fu viscera in CMT might be a space channel structure continuously consisted of unique molecules cavity that was imprinted with the supramolecularly template inside and outside of cells, through which the molecules in CMM interacted with the meridian and Zang-fu viscera. When small molecules in human body imprinted with macromolecules in meridian and Zang-fu viscera, in other words, they migrated along within imprinting channels of meridian and Zang-fu viscera on behavior of "Qi chromatography" impulsed by the heart beat, finally showed up on macroscopic the anisotropy of tissue and organ, as described namely as visceral manifestation in Chinese medical science. When small molecules in CMM interacted with imprinting channel on meridian and Zang-fu viscera, the natural properties and efficacy regularities of CMM was reflected on macroscopic. Therefore, the special representation forms of basic CMT is based on the macroscopic expression of "Qi chromatography" abided by imprinting effect regularities, and on whether the imprinted template of small molecules matched with cavity template of macromolecules in meridian and Zang-fu viscera, only is the adequate representation of supramolecular chemistry for them. The CMM materials is the mixture including single molecules and supramolecules. The compatibility for CM prescriptions can significantly change the function rules. Therefore in the study of basic CMT, we should pay special attention to the laws of supramolecular chemistry. It is the most essential differences of the CMT from the modern medicine which established by the laws of single molecular theories.

The relationship between partial disc displacement and mandibular dysplasia in female adolescents.

BACKGROUND: The objective of the present study was to investigate the possible relationship between partial disc displacement of the temporomandibular joint (TMJ) and mandibular dysplasia. MATERIAL/METHODS: The TMJ disc position of 46 female adolescents, with Class II, Division 1 malocclusion, aged 10.1 approximately 12.8 years, was quantitatively measured with magnetic resonance imaging (MRI) examinations. Meanwhile, mandibular size and form were determined from cephalometric radiographs. The possible correlation between disc status and mandibular morphology was assessed by means of the Pearson Correlation. In addition, the differences in mandibular size and form between the partial disc displacement group (displaced group) and the disc normal group were also assessed. RESULTS: The results of Pearson Correlation showed that the displaced group had shorter mandibular body length and total mandibular length (Go-Po/Ca-Dm, r=-0.500; Go-Po/ Dp-Cc/Cs-Cc, r=-0.500; Ar-Me/ Ca-Dm, r=-0.459). The patients with horizontal growing style presented more anterior displaced disc (N-S-Gn/ Ca-Dm, r=-0.492). Furthermore, a significantly shorter mandibular body length (Go-Po), a steeper mandibular plane (MP/FH) and a more obtuse mandibular angle (Ar-Go-Me) were found in the displaced group. However, no relationship between the size of the vertical mandible and the disc status was noticed. CONCLUSIONS: Partial disc displacement in Class II, Division 1 cases may be associated with the mandible with horizontal deficits instead of the mandible with vertical deficits.

Inhibition of alpha-mannosidase Man2c1 gene expression suppresses growth of esophageal carcinoma cells through mitotic arrest and apoptosis.

To study the effects of suppressed alpha-mannosidase Man2c1 gene expression on EC9706 human esophageal carcinoma cells, the cells were treated with short interfering RNA. Growth inhibition of EC9706 cells was observed when Man2c1 expression was inhibited in this way. Flow cytometric analysis showed accumulation of cells in S and G(2)-M phases, as well as cell apoptosis. The mitotic index test showed cell-cycle arrest at the M checkpoint. Although the percentage of cells in (pro)metaphase increased, the proportion of cells in anaphase and telophase decreased. Apoptosis was trigged by mitotic arrest. Furthermore, microtubules in EC9607 cells were examined by means of fluorescence staining of alpha-tubulin. Although control cells showed a nest-like microtubule network, the microtubule network in experimental cells was vague and condensed at the perinuclear region. Some cells with Man2c1 suppression had large protrusions of cytoplasm, some of which linked with the main body through a long, thin connection. Western blotting showed that tubulin polymerization was inhibited. The data imply that induction of mitotic arrest and consequent apoptosis resulted from microtubule disorganization, which appears to be one of the major cellular mechanisms by which suppressed expression of the Man2c1 gene causes growth inhibition of EC9706 esophageal carcinoma cells. In addition, Man2c1 suppression results in upregulation of E-cadherin, alpha-catenin, and beta-catenin expression in cells.

[Experimental BCSC-1 gene therapy on nasopharyngeal carcinoma mediated by adenovirus].

OBJECTIVE: To explore the therapeutic value of BCSC-1 in tumor gene therapy. METHODS: Recombinant adenovirus Ad5-BCSC-1 was prepared. Cell proliferation was assayed using CellTiter 96 Aqueous one solution cell proliferation assay kit. Ad5-BCSC-1 was injected into tumor with Ad5-egfp or with PBS injection as controls. The injections were repeated one weak later. After another 2 weeks, the mice were sacrificed and the tumors were excised and weighed. RESULTS: The growth of the CNE-2L2 cell infected with Ad5-BCSC-1 in vitro was remarkably slower than that of the controls, the wild type cell and the cell infected with Ad5-egfp. In contrast to the controls, the cells infected with Ad5-BCSC-1 aggregated and formed huge messes in the culture. The average weight of the CNE-2L2 tumors in mice was (2.28 +/- 0.73), (2.07 +/- 0.40), and (0.58 +/- 0.32) g for the tumors injected with PBS, Ad5-egfp, and Ad5-BCSC-1, respectively (Ad5-BCSC-1 vs. PBS or Ad5-egfp, P<0.05). CONCLUSION: Intra-tumor injection of Ad5-BCSC-1 can suppress the growth of CNE-2L2 tumor in nude mice, suggesting that BCSC-1 gene therapy may be effective for tumors with low or no expression of BCSC-1 gene.

Application of TQSM polypharmacokinetics and its similarity approach to ascertain Q-marker by analyses of transitivity in vivo of five candidates in Buyanghuanwu injection.

BACKGROUND: It is well-known that the public still have been facing on a severe issue about the inconsistency of quality and therapeutic efficacy of traditional medicines. Recently, Professor Chang-Xiao Liu has created a new promising concept for identifying relevant quality-markers (Q-marker) from herbs, their formulas and manufacturing products. Therefore, building up a new approach is necessary for us to bridge over quality to efficacy of pharmaceutical products. STUDY DESIGN: In this paper, five candidate Q-markers, astragaloside IV, paeonflorin, amygdalin, tetramethylpyrazine, ferulic acid in Buyanghuanwu injection (BYHWI) had been designed to carry out in rat by using single and polypharmacokinetic models for total quanta to ascertain adequate Q-marker. METHODS: The Q-marker transitivity in vivo was studied with polypharmacokinetic model and its similarity approach, which were modeled with TQSM principle. The Q-marker was ascertained with transitive similarity and bioavailability in polypharmacokinetics. Their concentrations in plasma sample of white rat were determined by RP-HPLC. Data analyses were used by the DAS software for singles and myself-written-program with EXCEL for multiples. RESULTS: In BYHWI, five candidate Q-marker pharmacokinetic profiles were singly fixed to two compartmental models in rat using classical compartmental analysis, but there were tremendous differences among which the candidate parameters were fluctuated from nearly 3552 folds to equivalency. The theoretical value of TQSM polypharmacokinetic parameters such as AUCT, MRTT, VRTT, CLT, VT over the mixure of five drugs were 110.8 ±â€¯51.91 mg min ml-1, 176.0 ±â€¯36.5 min, 39,921 ±â€¯4311 min2, 0.3116 ±â€¯0.02347 ml min-1 kg-1, 54.83 ±â€¯7.683 ml kg-1 respectively. The TQSM polypharmacokinetic parameters in astragaloside Ⅳ ordered by AUCT, MRTT, VRTT, CLT, VT were 110.8 ±â€¯51.91 mg min ml-1, 176.0 ±â€¯36.5 min, 39,921 ±â€¯4311 min2, 0.3116 ±â€¯0.02347 ml min-1 kg-1, 54.83 ±â€¯7.683 ml kg-1, respectively, which were closed to the theoretical values. TQSM similarity versus astragaloside Ⅳ was 0.9661. CONCLUSION: The results represented that the optimum Q-marker in BYHWI is astragaloside Ⅳ, whose transitivity in vivo similarity was close to the behavior of polypharmacokinetics with maximum bioavailability to the total quanta. It is feasible for Q-marker in CMMs to screen on the comparison of single pharmacokinetic behavior and bioavailability to the total quanta.

Novel mathematic models for quantitative transitivity of quality-markers in extraction process of the Buyanghuanwu decoction.

BACKGROUND: Nowadays, to research and formulate an efficiency extraction system for Chinese herbal medicine, scientists have always been facing a great challenge for quality management, so that the transitivity of Q-markers in quantitative analysis of TCM was proposed by Prof. Liu recently. In order to improve the quality of extraction from raw medicinal materials for clinical preparations, a series of integrated mathematic models for transitivity of Q-markers in quantitative analysis of TCM were established. Buyanghuanwu decoction (BYHWD) was a commonly TCMs prescription, which was used to prevent and treat the ischemic heart and brain diseases. In this paper, we selected BYHWD as an extraction experimental subject to study the quantitative transitivity of TCM. STUDY DESIGN: Based on theory of Fick's Rule and Noyes-Whitney equation, novel kinetic models were established for extraction of active components. Meanwhile, fitting out kinetic equations of extracted models and then calculating the inherent parameters in material piece and Q-marker quantitative transfer coefficients, which were considered as indexes to evaluate transitivity of Q-markers in quantitative analysis of the extraction process of BYHWD. METHODS: HPLC was applied to screen and analyze the potential Q-markers in the extraction process. Fick's Rule and Noyes-Whitney equation were adopted for mathematically modeling extraction process. Kinetic parameters were fitted and calculated by the Statistical Program for Social Sciences 20.0 software. The transferable efficiency was described and evaluated by potential Q-markers transfer trajectory via transitivity availability AUC, extraction ratio P, and decomposition ratio D respectively. The Q-marker was identified with AUC, P, D. RESULTS: Astragaloside IV, laetrile, paeoniflorin, and ferulic acid were studied as potential Q-markers from BYHWD. The relative technologic parameters were presented by mathematic models, which could adequately illustrate the inherent properties of raw materials preparation and affection of Q-markers transitivity in equilibrium processing. AUC, P, D for potential Q-markers of AST-IV, laetrile, paeoniflorin, and FA were obtained, with the results of 289.9 mAu s, 46.24%, 22.35%; 1730 mAu s, 84.48%, 1.963%; 5600 mAu s, 70.22%, 0.4752%; 7810 mAu s, 24.29%, 4.235%, respectively. CONCLUSION: The results showed that the suitable Q-markers were laetrile and paeoniflorin in our study, which exhibited acceptable traceability and transitivity in the extraction process of TCMs. Therefore, these novel mathematic models might be developed as a new standard to control TCMs quality process from raw medicinal materials to product manufacturing.

[Reduction of CD44 expression results in growth inhibition of human nasopharyngeal carcinoma cell CNE-2L2 in vitro].

OBJECTIVE: To study the effect of the inhibition of CD44 gene expression on the growth of human nasopharyngeal carcinoma cell CNE-2L2 in vitro. METHODS: CD44 gene expression in cells was suppressed by siRNA which was introduced into cells through retrovirus infection. Integration of siRNA into genomic DNA was examined by genomic PCR. CD44 gene expression in cells was detected by Western blot analysis. Cell growth in vitro was assayed using Cell Titer 96 AQueous One Solution Cell Proliferation Assay kit Promega. Cells were stained with propidium iodium and cell DNA content was detected upon a flow cytometer. RESULTS: siRNA was integrated into genomic DNA of host cells. The 4 cell pools integrated with one of the 4 siCD44s showed a significant inhibition of CD44 gene expression comparing to the controls, the wild type cell and the cell pool integrated with siegfp. The cell pools integrated with siCD44-1 or siCD44-2 showed the most profound inhibition. Growth of these 2 cells in vitro was compared to that of the controls and was found to be significantly inhibited. Cell DNA content analysis indicated 44.4%, 45.5%, 53.9%, and 53.3% in G0/G1 phase; 39.3%, 40.0%, 27.1%, and 28.2% in S phase; and 16.3%, 14.5%, 19.0%, and 18.5% in G2/M phase for the wild type cell, the cell pool integrated with siegfp, the cell pools integrated with siCD44-1, and the cell pools integrated with siCD44-2, respectively. CONCLUSION: Reduction in CD44 expression inhibit the growth of CNE-2L2 cell and affects the development of cells from G0/G1 into S phase, but may somehow promote cells to develop from S into G2/M phase.

[Ectopic expression of BCSC-1 gene results in enhancement of adhesion and cell cycling blockade of nasopharyngeal carcinoma CNE-2L2 cell].

OBJECTIVE: To study mechanisms of reduction of the malignant activities of human naso-pharyngeal carcinoma cell CNE-2L2 induced by ectopic expression of BCSC-1 gene. METHODS: DNA was stained with propidium iodide and assayed upon a flow cytometer. Chromosomes were stained with Hoechest 33258. Adhesion of CNE-2L2 cells was detected by cell aggregation test. Protein expression on CNE-2L2 cells was examined by Western blot. RESULTS: Cell cycle analysis showed that the percentage of CNE-2L2 cells was 55.1%, 43.4%, and 39.4% in G0/G1 phase, 25.2%, 28.7%, and 30.9% in S phase, and 19.7%, 27.9%, and 29.7% in G2/M phase for the cell with ectopic expression of BCSC-1 gene, wild type cell (W cells), and the cell transduced with the mock (M cell). Many mitotic cells were found in W cells and M cells. In contrast, almost no mitotic cell was observed in the cells with ectopic expression of BCSC-1 gene. Ectopic BCSC-1 expression resulted in cell aggregation, enhanced expression of E-cadherin, cx-catenin, and p53. CONCLUSIONS: Ectopic BCSC-1 expression causes enhancement of adhesion of CNE-2L2 cells associated with enhanced expression of E-cadherin and alpha-catenin, arrest of cell in G1 phase, which may be associated with enhanced expression of p53. These alteration may play a role in the reduction of malignant activities of the cells with ectopic expression of BCSC-1 gene.

[Inhibition of malignant activities of nasopharyngeal carcinoma cell by ectopic expression of BCSC-1 gene].

OBJECTIVE: To study effects of ectopic expression of BCSC-1 gene on the malignant activi-BCSC-1 cDNA was isolated by RT-PCR ties of human nasopharyngeal carcinoma cell CNE-2L2. METHODS: and inserted into pMAL-c2X and pcDNA4/myc-His A. BCSC-1 protein was expressed in prokaryocytes. Rabbit antiserum to BCSC-1 was developed by means of immunization of rabbit with the BCSC-1 protein. Expression of BCSC-1 gene in wild type CNE-2L2 cell (W cell) was examined by real-time RT-PCR and immunofluorescence staining with the antiserum as a probe. pcDNA4/myc-His A-BCSC-1 was transfected into W cell at the presence of LipofectAmine. The cells were selected by G418 and cloned. Ectopic expression of BCSC-1 gene in W cell was examined by Western blot. Cell growth was detected by drawing of growth curves and colony formation tests. Cells were inoculated into nude mice. Size of tumors was assayed once a week. Lungs of the mice were sectioned continuously and metastatic loci in lungs were examined upon a microscope. RESULTS: Rabbit BCSC-1 antiserum was prepared. Expression of BCSC-1 gene in W cell was found to be very low. CNE-2L2 cell with ectopic expression of BCSC-1 gene was developed. Growth in vitro, colony formation, tumorigenesis in nude mice, and lung metastasis of the tumor were profoundly inhibited of the cell with ectopic expression of BCSC-1 gene in comparison with controls, wild type cell and the cell transfected with mock. Conclusion Ectopic expression of BCSC-1 gene exerts profound inhibitive effect on the malignant activities of CNE-2L2 cell.

[Experimental siCD44-targeted therapy of human nasopharyngeal carcinoma mediated by adenovirus].

OBJECTIVE: To explore the possibility of treating solid tumor with siCD44. METHODS: Human nasopharyngeal carcinoma cell CNE-2L2 with high expression of CD44 was used in this study. The malignant activities of cells were examined by colony formation test, tumorigenesis, and lung metastasis of the tumor in nude mice. Ad5-siCD44 was constructed and adenoviruses were produced in 293 cells. CNE-2L2 cells were subcutaneously inoculated into nude mice. When tumors grew to 50-100 mm3, Ad5-siCD44 was injected into tumors, and Ad5-egfp and PBS were also injected as controls. The size and weight of tumors were compared after 2 weeks. RESULTS: Suppression of CD44 expression profoundly inhibited the malignant activities of CNE-2L2 cell. The average sizes of the tumors were (3.139 +/- 0.850), (3.612 +/- 0.888), and (1.512 +/- 0.742) cm3 after the intra-tumor injection of PBS, Ad5-egfp, and Ad5-siCD44, respectively, after two weeks. Significant difference was found between Ad5-siCD44 group and control groups (P < 0.05). The average weights were (2.28 +/- 0.73), (1.83 +/- 0.26), and (1.20 +/- 0.64) g, respectively, and significant difference was also found between Ad5-siCD44 group and control groups (P < 0.05). CONCLUSION: Intra-tumor injection of Ad5-siCD44 can exhibit the therapeutic effect on the tumor inoculated with CNE-2L2 cells with high expression of CD44 in nude mice.