Histone methyltransferase SETDB1 promotes colorectal cancer proliferation through the STAT1-CCND1/CDK6 axis.

Upregulation of histone methyltransferase SET domain bifurcated 1 (SETDB1) is associated with poor prognosis in cancer patients. However, the mechanism of oncogenicity of SETDB1 in cancer is hitherto unknown. Here, we show that SETDB1 is upregulated in human colorectal cancer (CRC) where its level correlates with poor clinical outcome. Ectopic SETDB1 promotes CRC cell proliferation, whereas SETDB1 attenuation inhibits this process. Flow cytometry reveals that SETDB1 promotes proliferation by driving the CRC cell cycle from G0/G1 phase to S phase. Mechanistically, SETDB1 binds directly to the STAT1 promoter region resulting in increased STAT1 expression. Functional characterization reveals that STAT1-CCND1/CDK6 axis is a downstream effector of SETDB1-mediated CRC cell proliferation. Furthermore, SETDB1 upregulation is sufficient to accelerate in vivo proliferation in xenograft animal model. Taken together, our results provide insight into the upregulation of SETDB1 within CRC and can lead to novel treatment strategies targeting this cell proliferation-promoting gene.

[Study on formation and changes of Dao-di herbs production origin of Gastrodia elata].

Gastrodia elata has been used in China for more than 2 000 years and it is a kind of valuable traditional Chinese medicine. The originrecords of G. elata were Mount Tai of Shandong and and Mount Song of Henan, which began in Wupu Bencao of Wei Jin Dynasties, and Tai'an and its surrounding areas had been the Do-di herbs production areas. But from the beginning of the Republic of China, G. elata origin has undergone major changes, Do-di herbs production areas moved westward to the southwest.In this paper,through literature research and field visits, we studied the formation and changes of Do-di herbs production areas of G. elata. The cultivation history and current main producing area of G. elata was also introduced. On this basis, we profoundly summarized the reasons of Do-di herbs production areas formation and changes from the nature, society, transportation, humanities and germplasm resources.Combining the ancient herbal medicine and the characteristics of modern producing areas, the planting strength of G. elata could be strengthened in the hope of providing reference for the quality evaluation and cultivation of G. elata.

Low Temperature Soda-Oxygen Pulping of Bagasse.

Wood shortages, environmental pollution and high energy consumption remain major obstacles hindering the development of today's pulp and paper industry. Energy-saving and environmental friendly pulping processes are still needed, especially for non-woody materials. In this study, soda-oxygen pulping of bagasse was investigated and a successful soda-oxygen pulping process for bagasse at 100 °C was established. The pulping parameters of choice were under active alkali charge of 23%, maximum cooking temperature 100 °C, time hold at maximum temperature 180 min, initial pressure of oxygen 0.6 MPa, MgSO4 charge 0.5%, and de-pithed bagasse consistency 12%. Properties of the resultant pulp were screened yield 60.9%, Kappa number 14, viscosity 766 dm³/kg, and brightness 63.7% ISO. Similar pulps were also obtained at 110 °C or 105 °C with a cooking time of 90 min. Compared with pulps obtained at higher temperatures (115-125 °C), this pulp had higher screened yield, brightness, and acceptable viscosity, while the delignification degree was moderate. These results indicated that soda-oxygen pulping at 100 °C, the lowest cooking temperature reported so far for soda-oxygen pulping, is a suitable process for making chemical pulp from bagasse. Pulping at lower temperature and using oxygen make it an environmental friendly and energy-saving pulping process.

[Induction and analysis for NIR features of frequently-used mineral traditional Chinese medicines].

In order to provide theoretical basis for the rapid identification of mineral traditional Chinese medicines(TCM) with near infrared (NIR)diffuse reflectance spectroscopy, Characteristic NIR spectra of 51 kinds of mineral TCMs were generalized and compared on the basis of the previous research, and the characteristic spectral bands were determined and analyzed by referring to mineralogical and geological literatures. It turned out that the NIR features of mineral TCMs were mainly at 8 000-4 000 cm ⁻¹ wavebands, which can be assigned as the absorption of water, -OH and[CO3 ²â»] and so on. Absorption peaks of water has regularity as follows, the structure water and -OH had a combined peak which was strong and keen-edged around 7 000 cm ⁻¹, the crystal water had two strong peak around 7 000 cm ⁻¹ and 5 100 cm ⁻¹, and water only has a broad peak around 5 100 cm ⁻¹. Due to the differences in the crystal form and the contents of water in mineral TCMs, NIR features of water in mineral TCMs which could be used for identification were different. Mineral TCMs containing sulfate are rich in crystal water, mineral TCMs containing silicate generally had structure water, and mineral TCMs containing carbonate merely had a little of water, so it was reasonable for the use of NIR spectroscopy to classify mineral TCMs with anionic type. In addition, because of the differences in cationic type, impurities, crystal form and crystallinity, mineral TCMs have exclusive NIR features at 4 600-4 000 cm ⁻¹, which can be assigned as Al-OH, Mg-OH, Fe-OH, Si-OH,[CO3 ²â»] and so on. Calcined mineral TCMs are often associated with water and main composition changes, also changes of the NIR features, which could be used for the monitoring of the processing, and to provide references for the quality control of mineral TCMs. The adaptability and limitation of NIR analysis for mineral TCMs were also discussed:the majority of mineral TCMs had noteworthy NIR features which could be used for the NIR analysis. And the NIR features of a few mineral TCMs were inapparent, such as Fluoritum, Realgar and Cinnabar, for which the Raman spectroscopy can be adopted alternatively.

[Analysis on 24 Samples of Fluoritum by XRD Pattern].

Objective: To provide the reference for effective identification of Fluoritum,by using X-ray diffraction technique to analyze Fluoritum samples which had different morphological features. Methods: According to the China Pharmacopeia( 2010 edition),the24 samples of commercial Fluoritum were identified and their contents of Ca F2 were determined. XRD technique was applied to analyze phase compositions and content from Fluoritum samples, to ensure quality, and to summarize the correlation between traits and quality. Results: Sample 1 ~ 7 and 13 were Fluoritum, samples 8 ~ 12 were inferior products which were doped, and sample 14 ~ 24 were counterfeit products. Fluorite was the main phase of Fluoritum, and often accompanying a small amount of quartz. Phase compositions of counterfeit Fluoritum whose impurity content were high were relatively complicated, and the contents of Ca F2 were far below the standards value of the China Pharmacopeia( 2010 edition). Fluoritum were easy to be shattered into tiny sand which were green or purple, hyaline and lustered, with color becoming shallow. Conclusion: By picking to remove impurity, inferior products can be used for medicine. Because impurity content are high and the impurities are difficult to be separated, counterfeit products can not be used for medicine. Characteristics of powder can be used for supplement the identification for Fluoritum. And using XRD technique can accurately identify Fluoritum samples which have different morphological features.

[Identification of Mineral Medicine of Chloriti Lapis,Micae Lapis Aureus and Vermiculitum by X-ray Diffraction].

Objective: To analyze the mineral medicine of Chloriti Lapis, Micae Lapis Aureus and Vermiculitum by X-ray diffraction, then to guide the identification and quality assessment of them. Methods: XRD Fourier patterns were collected from powder samples to analyze phase compositions, and to determine the original mineral resources of Chloriti Lapis, Micae Lapis Aureus and Vermiculitum by comparing with their characteristic traits. First derivative + vector normalization and 21 point smoothing were used to pretreat the selected spectrum band from 0. 68 ~ 1. 77 nm. Then the data were analyzed by fuzzy cluster. Results: It was found that the original mineral resource of seven powder samples of Chloriti Lapis was biotite schist belonging to metamorphic mineral. The original mineral resource of three powder samples of high-quality Micae Lapis Aureus was vermiculite biotite schist belonging to metamorphic mineral. The original mineral resources of three powder samples of Vermiculitum were phlogopite and vermiculite phlogopite. Conclusion: The method of X-ray diffraction analysis is accurate and rapid, which can be used for the identification and quality evaluation of Chloriti Lapis, Micae Lapis Aureus and Vermiculitum.

[Taxonomy and Distribution of Chinese Medicinal Centipedes].

Objective: To study the taxonomy and distribution of Chinese medicinal centipedes. Methods: The species of Chinese medicinal centipedes were investigated in the light of their morphology. According to the feature of life, the distribution of centipedes were explored. Results: There were 12 centipede species in China, and seven of them were used for medical, the species could be effectively distinguished by the identification key. It was suggested that their characteristics were related to the climatic factors such as temperature, humidity, altitude and air pressure. Conclusion: The distribution of medicinal centipede is characteristics of "three river system distribution belts" and "three geographical distribution areas". The results provide the basis for the development and application of medicinal centipedes in China.

[Study on identification of traditional Chinese medicine Yangqishi and Yinqishi by X-ray diffraction].

The aim of this paper is to clarify the mineral origin of traditional Chinese medicine (TCM) Yangqishi and Yinqishi and guide identification of the both, by X-ray diffraction (XRD) Fourier patterns. Morphological identification and conventional physical and chemical analysis wee used to identify 22 batches of Yangqishi and Yinqishi. It used XRD Fourier patterns which has been collected from sample powders to analyze phase composition. It has been found experimentally that the mineral origin of Yinqishi is Talc schist and the mineral origin of Yangqishi is tremolite and actinolite. The results also showed that the method using XRD can get not only an accurate but also rapid identification of Yangqishi and Yinqishi. There are many differences in medicinal properties, efficacy, indications and composition of Yangqishi and Yinqishi, so be careful not to mix them up.

[Quantitative models of Raman spectroscopy for five kinds of traditional Chinese medicine containing CaCO3 based on an improved siPLS].

The aim of this paper is to apply Raman spectroscopy technique to develop rapid quantitative models for five kinds of Traditional Chinese Medicine containing CaCO3. In the experiment, Raman spectras of 67 batch of sample including Otolithum Sciaenae, Galaxeae Os, Ophicalcitum, Calcite, Stalactite and their mixture which had different content of CaCO3 were collected, and the quantitative models were established by using an improved siPLS to optimize the characteristic spectral bands and using the CaCO3 contents which were measured by EDTA titration method as references. Compared with the results by EDTA titration, the established quantitative model for CaCO, content showed a prediction result that the average relative deviation of the prediction results is 2. 71% and the average recovery rate was 100.46%, when the content is between 0.465 4-0.999 7, and when the characteristic spectral bands of 1 290-1 280, 730-714, 700-690, 660-650, 465-460, 455-445, 405-385 cm(-1) had been optimized. The result also showed that the model using Raman spectroscopy and based on an improved siPLS can get a rapid determination for contents of 5 kinds of Traditional Chinese Medicine containing CaCO3.

[Development history of methodology of Chinese medicines' authentication].

This paper reviewed the emergence process of the subject and methodology of Chinese Medicines' Authentication. Based on the research progress and major achievements acquired in research of each methodology including identification of origin, description, microscopic, physical, chemical and biological characteristics of Chinese medicines, it is expounded that the development process of each methodology combined modem digital technology, information science and its own characteristics. And the development direction is further described for methodology of Chinese Medicines' Authentication towards systematization and informationization.

[Molecular identification of Hibiscus syriacus and its adulterants using ITS2 barcode].

OBJECTIVE: To identify Hibiscus syriacus and its adulterants using DNA barcoding technique. METHODS: Nine samples of five species were PCR amplified and sequenced, and twelve samples were downloaded from the GenBank. The intra-specific and interspecific K2P distances were calculated, and neighbor-joining( NJ) tree was constructed by MEGA 5.0. RESULTS: The results showed the intra-specific genetic distances of Hibiscus syriacus were ranged from 0.009 to 0.056, which were far lower than inter-specific genetic distances between Hibiscus syriacus and its adulterants (0.236 - 0.301). Variable sites within Hibiscus syriacus ranged from 2 to 9 which were far less than the adulterants (45 - 52); Different samples of Hibiscus syriacus were gathered together and could be distinguished from its adulterants by NJ tree. CONCLUSION: ITS2 can discriminate Hibiscus syriacus from its adulterants correctly. The ITS2 region is an efficient barcode for authentication of Hibiscus syriacus and its adulterants.

[A NIR qualitative and quantitative model of 8 kinds of carbonate-containing mineral Chinese medicines].

The aim of this paper is to apply near infrared spectroscopy techniques to construct a rapid identification method for 8 kinds of mineral Chinese Medicines containing carbonates. The qualitative model using clustering analysis method in OPUS software can identify accurately 8 kinds of carbonate-containing mineral Chinese medicines. The near-infrared quantitative model was established by using partial least squares method (PLS) for 7 mineral Chinese Medicines in which main component is calcium carbonate. Compared with the results by EDTA titration, the established quantitative analysis model for calcium carbonate content showed a good prediction result that when the content is between 47.61% -99.17%, the average relative deviation of the prediction result is 0.24% and the average recovery rate was 100.3%. The results also showed that the model using near infrared spectroscopy can get not only a rapid identification of the 8 mineral Chinese medicines containing carbonates, but also an accurate and reliabe content determination of calcium carbonate for the 7 mineral Chinese medicines which contain the component.

[Influence of different processed methods on 10 kinds of ginsensides in Panacis Quinquefolii Radix].

The influence on 10 kinds of ginsensides of different processed methods of Panacis Quinquefolii Radix was discussed. White Panacis Quinquefolii Radix (sliced and dried at -80 °C), red Panacis Quinquefolii Radix( steamed, sliced and dried at -80 °C) and commercial Radix Panacis Quinquefolii (dried by electric blast air) processed by different methods. HPLC-PDA-ESI- MS method was established before by our team. Ten kinds of ginsenosides of them were determined. The content of total ginsenosides were as follow: commercial Panacis Quinquefolii Radix > white Panacis Quinquefolii Radix > red Panacis Quinquefolii Radix. Compared with white Panacis Quinquefolii Radix, the content of Re, Rc, Rb3 and Rb2 of Red Radix Panacis Quinquefolii decreased but increased that of Rg,, Rb1. Both Rg2 and Rg, were not found in white Panacis Quinquefolii Radix and commercial Panacis Quinquefolii Radix by PDA detector, and low response in ESI-MS, while red Panacis Quinquefolii Radix was to the high content that of 0. 027% and 0.040 1%. The constituent of RA0 of red Panacis Quinquefolii Radix was higher than the other two. After Panacis Quinquefolii Radix processed, the kind and content of ginsensides were significantly changed. The constituent of some kinds of ginsensides was increased and some decreased. Rf was not found in all Panacis Quinquefolii Radix samples which were consistent with the former documents.

[Study on quality standard of Selaginella moellendorffii].

OBJECTIVE: To study the quality standard of Selaginella moellendorffii. METHODS: The macroscopic and microscopic characteristics were observed, amentoflavone was used as reference substance in the TLC identification and HPLC method was used to determine the content of amentoflavone. RESULTS: The TLC method used GF254 taking toluene-ethyl formate-formic acid (5: 4: 0. 5) as the developer and ethanol solution of ferric chloride as the chromogenic reagent with the results that it appeared four clear spots. The HPLC method took amentoflavone as the reference substance, and acetonitrile-water (containing 3% tetrahydrofuran and 3% trifluoroacetic acid) as the mobile phase; Detection wavelength was 330 nm. The contents of samples from 3 different batches were determined, and the lowest content of amentoflavone was 0.4%. CONCLUSION: This method is reliable and accurate, which can be used for the quality control of Selaginella moellendorfii.

[Identification of Peucedani Radix and its adulterants by DNA barcoding technique].

OBJECTIVE: To identify Peucedani Radix and its adulterants using DNA barcoding technique. METHODS: Total genomic DNA was isolated from Peucedani Radix and its adulterants. Nuclear DNA ITS2 sequences were amplified and purified PCR products were sequenced. Sequence assembly and consensus sequence generation were performed using the CodonCode Aligner V3.0. The Kimura 2-Parameter(K2P) distances were calculated using software MEGA 4. 0. Identification analyses were performed using BLAST1, Nearest Distance and Neighbor-Joining (NJ) methods, and the secondary structure of the ITS2 sequence differences between species were analyzed. RESULTS: Different samples of Peucedani Radix were gathered together and distinguished from its adulterants by NJ tree. The ITS2 secondary structure showed that Peucedani Radix could be differentiated obviously from its adulterants. CONCLUSION: ITS2 sequence is able to identify Peucedani Radix and its adulterants correctly, which provides a scientific basis for fast and accurate identification of the herb.

[A new monoterpenoid glucoside from the twigs of Chamaecyparis obtusa var. breviramea f. crippsii].

To investigate the chemical constituents of Chamaecyparis obtusa var. breviramea f. crippsii, various column chromatography and spectroscopic methods were used for the isolation and elucidation of compounds. One new monoterpenoid glucoside, (4S)-4-isopropylcyclohex-l-enecarboxylic acid 4-O-beta-D-glucopyranoside (1), together with five known compounds, (4R)-p-menth-l-ene-7, 8-diol 7-O-beta-D-glucopyranoside (2), skimmin (3), 7-[[6-O-(6-deoxy-alpha-L-mannopyranosyl)-beta-D-glucopyranosyl]oxy]-2H-1-benzopyran-2-one (4), stigmast-4-en-3-one (5) and 1, 4-benzenedicarboxylic acid 1-butyl-4-(2-methylpropyl) ester (6) were isolated and identified from the twigs of this plant. All compounds were isolated from this plant for the first time. The methanol extract of this plant showed cytotoxicity on cancer cell lines A549, BGC-823, Du145 and MDA-MB-231 with IC50 values of 0.94, 1.07, 0.95 and 0.96 microg x mL(-1), respectively. Yet, compounds 1, 2 and 3 showed no cytotoxicity on cancer cell lines HeLa, BGC-823 and A549.

[Chemical constituents of ethyl acetate extract from Polygonum perfoliatum].

OBJECTIVE: To study the chemical constituents of Polygonum perfoliatum. METHODS: The chemical constituents of Polygonum perfoliatum were isolated and purified by chromatography on silica gel, Sephadex LH-20 and Semi-preparative HPLC. The structures of compounds were elucidated and identified by physicochemical properties, H-NMR, 13C-NMR and ESI-MS. RESULTS: Ten compounds were isolated and identified as 5-hydroxymethyl-2-furaldEhyde (1), methyl caffeoate (2), protocatechuic aldehyde (3), quercetin (4), pinocernbrin (5), catechin (6), taxifolin (7), taxifolin-3-O-beta-D-xylopyranoside (8), 13-epitorulosal (9), coumarin-7-O-beta-D-glucose glycosidic (10). CONCLUSION: [corrected] Compounds 6-9 are isolated from this plant for the first time.

[Study on the pollen viability and stigma receptivity of Chrysanthemum morifolium 'Fubaiju'].

OBJECTIVE: To study the blooming habits, pollen viability and stigma receptivity of Chrysanthemum morifolium and provide theoretical basis for its breeding. METHODS: Explored the blooming habits by dynamic observation on the process of blossom, evaluated the pollen viability by germination in vitro culture method and estimated stigma receptivity by benzidine-hydrogen peroxide method. RESULTS: About the pollen viability, there were no significant differences between the flowers which in the same round of the capitulum; Tubular flowers in the center of a capitulum were significantly higher than that on the edge; In the morning pollen vitality gradually raised, during 11: 00 - 14: 00 maintained the highest, and then gradually decreased; Tubular flower began to loose powder on the third day, during 4th - 6th day the pollen viability was highest, respectively was 35.12%, 39.89%, 38.12%, then gradually decreased, on the 15th day was only 7.41%, finally turned into wither. Regard to the stigma receptivity, the center of a capitulum were significantly higher than that on the edge, outer edge ligulate flower had no receptivity; Revealed the strongest during 13: 00 - 14:00 in one day; During the 5th - 7th day was the strongest after flowering. The regulation of the stigma secreted mucus existed great consistency with the stigma receptivity, namely the stigma receptivity usually was strong when it secreted large number mucus. CONCLUSION: Understand the blossom habits of Chrysanthemun morifolium, as well as the dynamic changes regulation of pollen viability and stigma receptivity during its blossom, which could be used to select the flowers in a capitulum which are on the more suitable period and position for artificial pollination and hybridization breeding research.

[Investigation on bupleurum resources in northwest area of Hubei Province].

OBJECTIVE: To investigate the species and the distribution of Bupleurum genus in northwest of Hubei province and provide basis for the standardization planting of Bupleurum in the region. METHODS: Investigated and collected specimens on the spot, com- pared them with literatures and specimen collected before. RESULTS: The main species of Bupleurum plants was B. marginatum, only very few of them was B. scorzonerifolium, no B. chinense was found in northwest of Hubei province. CONCLUSION: B. marginatum is the mainstream species of Bupleurum plants and the roots of B. marginatum are the actual source of commodity with the name of "Beichaihu (the roots of B. chinense)" in northwest of Hubei province.

Ethanol Extract from Ampelopsis sinica Root Exerts Anti-Hepatitis B Virus Activity via Inhibition of p53 Pathway In Vitro.

Ampelopsis sinica root is widely used in Chinese folk medicine for treating liver disorders caused by the hepatitis B virus (HBV). The present study was performed in order to investigate the anti-HBV activity and mechanisms of the ethanol extract from A. sinica root (EASR) in vitro. The antiviral activity of EASR was examined by detecting the levels of HBsAg, HBeAg and extracellular HBV DNAs in stable HBV-producing human hepatoblastoma HepG2 2.2.15 cells. We found that EASR effectively suppressed the secretion of HBsAg and HBeAg from HepG2 2.2.15 cells in a dose-dependent manner, and it also suppressed the amount of extracellular HBV DNA. After EASR treatment, the percentage of apoptotic cells was found to be significantly higher than that of control by flow cytometric analysis. A luciferase reporter gene assay was used to determine the effects of EASR on the activities of HBV promoters and intracellular signaling pathways. The results showed that EASR selectively inhibited the activities of HBV promoters (Cp, S1p and Fp) and the p53 signaling pathway in HepG2 cells significantly. These data indicate that EASR exerts anti-HBV effects via inhibition of HBV promoters and the p53-associated signaling pathway, which helps to elucidate the mechanism underlying the potential therapeutic value of EASR.