[Quality value transfer of substance benchmarks in Sanpian Decoction].

According to the polarity of different components in Sanpian Decoction, two fingerprints were established. Then the substance benchmark freeze-dried powder of 15 batches of Sanpian Decoction was prepared, followed by the determination of the fingerprints, index component content, and dry extract rates, the identification of attribution of characteristic peaks, and the calculation of similarities between these fingerprints and the reference(R), the content and transfer rate ranges of ferulic acid, sinapine thiocyanate, liquiritin, and glycyrrhizic acid, and the dry extract rate range. The results showed that the similarities of 15 batches of the substance benchmark fingerprints with R were all greater than 0.900.Further summarization of the characteristic peaks revealed that there were a total of 20 characteristic peaks in fingerprint 1, among which, eight were from Sinapis Semen, four from Paeoniae Radix Alba, six from Chuanxiong Rhizoma, and two from Glycyrrhizae Radix et Rhizoma. A total of 16 characteristic peaks were observed in fingerprint 2, including one from Sinapis Semen, three from Paeoniae Radix Alba, eight from Chuanxiong Rhizoma, and four from Glycyrrhizae Radix et Rhizoma. The average dry extract rate of 15 batches of substance benchmarks was 18.25%, with a dry extract rate range of 16.28%-20.76%. The index component content and transfer rate ranges were listed as follows: 0.15%-0.18% and 38.81%-58.05% for ferulic acid; 0.26%-0.42% and 36.51%-51.02% for sinapine thiocyanate; 0.09%-0.15% and 48.80%-76.61% for liquiritin; 0.13%-0.24% and 23.45%-35.61% for glycyrrhizic acid. The fingerprint, dry extract rate, and index component content determination was combined for analyzing the quality value transfer of substance benchmarks in the classic prescription Sanpian Decoction.The established quality evaluation method for the substance benchmarks was stable and feasible, which has provided a basis for the quality control of Sanpian Decoction and the follow-up development of related preparations.

[Application progress on taste evaluation methods for oral preparations].

Taste is an important factor affecting the medicinal properties of oral preparations and patient compliance with medication, and also an important evaluation index for oral preparation design and clinical application. How to characterize the taste objectively, accurately, simply, and efficiently is a bottleneck problem that restricts the taste design, development, and utilization of oral preparations. At present, the commonly used taste assessment methods for oral preparations are traditional human taste panel, electronic tongue, animal preference test, in vitro release study, and electrophysiological test. The traditional human taste panel is the first choice for taste evaluation, but it is limited by poor subjectivity and reproducibility. Therefore, despite some limitations, the other four taste assessment methods have been applied in the pharmaceutical industry as auxiliary methods. This study reviewed the detection principles, applicability, advantages, and disadvantages of the above methods to provide references for the taste correction research and taste assessment of oral preparations, improve patient compliance and the competitiveness of oral preparation products in the industry, and promote the development of oral preparation technologies.

Therapeutic effects of Chinese medicine Di-Long (Pheretima vulgaris) on rheumatoid arthritis through inhibiting NF-κB activation and regulating Th1/Th2 balance.

Chinese medicine Di-Long, the dried body of Pheretima vulgaris (Chen) has been used for the treatment of joint inflammation, arthralgia and numbness of limbs for many years. This study was to investigate the anti-rheumatoid arthritis (RA) effects of Di-Long and to explore its possible mechanisms. The identification and quantification of representative components in Di-Long extracts (DL) were carried out by HPLC analysis. The anti-RA effects and mechanisms of DL were studied in CIA mice, RAW 264.7 macrophages and spleen T lymphocytes. The Th1/Th2 cell ratio in CIA mice spleens were determined by Flow cytometry. The cytokine levels were determined by ELISA method. The expressions of p-NF-κB p65 in ankle joints of CIA mice were detected by Immunohistochemistry analysis. The phosphorylation of NF-κB signaling pathway in RAW 264.7 macrophages and expressions of T-bet and GATA-3 in CIA mice spleens were determined by Western blots. The treatment with DL significantly decreased the paw thickness, arthritis scores and inflammatory cells infiltration in CIA mice. The TNF-α, IL-6 concentrations in both mice serum and macrophages secretion were markedly reduced with the treatment of DL, as well as the phosphorylation of NF-κB pathway. DL inhibited the expressions of T-bet and GATA-3 and decreased Th1/Th2 cells ratio in CIA mice spleens. DL reduced IFN-γ, IL-2 levels in mice serum and spleen T lymphocytes, and increased IL-4 levels in CIA mice serum. Chinese medicine Di-Long have significant anti-RA effects. The mechanisms might be inhibiting the activation of NF-κB signaling pathway and regulating the balance of Th1/Th2 cells.

[Analysis on quality value transmitting of substance benchmark of Houpo Wenzhong Decoction].

By preparing 15 batches of lyophilized powder samples of substance benchmark in Houpo Wenzhong Decoction,the fingerprint,index component content and extract rate were determined,and the characteristic peaks,the range of similarity with the reference map,the content range and transfer rate range of magnolol,hesperidin,glycyrrhizic acid and pinocembrin,the extract rate range and the change range were clarified. The results showed that the similarity between the fingerprint of substance benchmark and the reference map R generated from the 15 batches of substance benchmark samples was higher than 0. 90. The assignment of the characteristic peaks in the full prescription's fingerprint of the herbs except Poria cocos was clarified. Nineteen characteristic peaks were assigned,and 12 characteristic peaks were assigned by the reference substance,of which 4 were from Magnolia ocinalis Cortex,5 from Exocarpium Citri Rubrum,2 from Radix aucklandiae,3 from Glycyrrhiza Radix et Rhizoma,4 from Semen Alpiniae Katsumadai,and one from Rhizoma Zingiberis and Zingiber officinale Roscoe. The index component content range and transfer rate range were 0. 80%-1. 14% and 20. 25%-39. 61% for hesperidin,0. 49%-0. 79% and 23. 09%-33. 87%for glycyrrhizic acid,0. 03%-0. 07% and 3. 55%-10. 09% for pinocembrin,0. 15%-0. 38% and 8. 08%-24. 35% for magnolol. The extract rate range and the change range were22. 60%-25. 57% and 12. 67%-23. 68% respectively. In this study,we introduced the concepts of index component content,fingerprint,extract rate,explored the transfer relation of quality value transmitting of substance benchmark in Houpo Wenzhong Decoction,and initially established the quality standard of Houpo Wenzhong Decoction,all of which would provide ideas for the development and research of similar prescriptions.

Isolation, purification, and identification of antialgal substances in green alga Ulva prolifera for antialgal activity against the common harmful red tide microalgae.

Ten compounds (1~10) were successfully isolated from green algae Ulva prolifera through the combination of silica gel column chromatography, Sephadex LH-20 column chromatography and repeated preparative thin-layer chromatography. These ten compounds showed antialgal activity against red tide microalgae. Among them, compounds 3, 6, and 7 showed stronger antialgal activity against red tide microalgae. Furthermore, their structure was identified on the basis of spectroscopic data. There are three glycoglycerolipids: 1-O-octadecanoic acid-3-O-ß-D-galactopyranosyl glycerol (2), 1-O-palmitoyl-3-O-ß-D-galactopyranosyl glycerol (4), and 1-O-palmitoyl-2-O-oleoyl-3-O-ß-D-galactopyranosyl glycerol (5); two monoglycerides: glycerol monopalmitate (1), 9-hexadecenoic acid, 2,3-dihydroxypropyl ester (3); two terpenoids: loliolide (6) and lsololiolide (7); one lipid-soluble pigments: zeaxanthin (8); one sterol: cholest-5-en-3-ol (9); and one alkaloid: pyrrolopiperazine-2,5-dione (10). These compounds were isolated from U. prolifera for the first time, and compounds 2, 3, 5, and 8 were isolated from marine macroalgae for the first time.

[Isolation, Purification and Identification of Antialgal Activity Substances of Ethyl Acetate Extracts from the Submerged Macrophytes Potamogeton crispus].

Previous studies showed that ethyl acetate extracts from the submerged macrophytes Potamogeton crispus can significantly inhibit the growth of Karenia mikimitoi. Further, two antialgal activity compounds (1-2) were successfully isolated from this submerged macrophytes through a combination of silica gel column chromagraphy and repeated preparative thin-layer chromatography in this paper. These two antialgal activity compounds exhibited antialgal active against Karenia mikimitoi. Furthermore, their structure were identified on the basis of spectroscopic data: one flavonid named Trichodermatides B, and one alkaloid named 2-methylheptylisonicotinate. These two compounds were for the first time isolated from both Potamogeton crispus and submerged macrophytes.

Physiological and biochemical responses of Ulva prolifera and Ulva linza to cadmium stress.

Responses of Ulva prolifera and Ulva linza to Cd(2+) stress were studied. We found that the relative growth rate (RGR), Fv/Fm, and actual photochemical efficiency of PSII (Yield) of two Ulvaspecies were decreased under Cd(2+) treatments, and these reductions were greater in U. prolifera than in U. linza. U. prolifera accumulated more cadmium than U. linza under Cd(2+) stress. While U. linza showed positive osmotic adjustment ability (OAA) at a wider Cd(2+) range than U. prolifera. U. linza had greater contents of N, P, Na(+), K(+), and amino acids than U. prolifera. A range of parameters (concentrations of cadmium, Ca(2+), N, P, K(+), Cl(-), free amino acids (FAAs), proline, organic acids and soluble protein, Fv/Fm, Yield, OAA, and K(+)/Na(+)) could be used to evaluate cadmium resistance in Ulva by correlation analysis. In accordance with the order of the absolute values of correlation coefficient, contents of Cd(2+) and K(+), Yield, proline content, Fv/Fm, FAA content, and OAA value of Ulva were more highly related to their adaptation to Cd(2+) than the other eight indices. Thus, U. linza has a better adaptation to Cd(2+) than U. prolifera, which was due mainly to higher nutrient content and stronger OAA and photosynthesis in U. linza.

[Growth inhibition of the four species of red tide microalgae by extracts from Enteromorpha prolifera extracted with the five solvents].

To study the effects of extracts of Enteromorpha prolifera on the growth of the four species of red tide microalgae (Amphidinium hoefleri, Karenia mikimitoi, Alexandrium tamarense and Skeletonema costatum), the extracts were extracted with five solvents (methanol, acetone, ethyl acetate, chloroform and petroleum ether), respectively. Based on the observation of algal morphology and the measurement of algal density, cell size and the contents of physiological indicators (chlorophyll, protein and polysaccharide), the results showed methanol extracts of E. prolifera had the strongest action. The inhibitory effects of A. hoefleri, K. mikimitoi, A. tamarense and S. costatum by the methanol extracts were 54.0%, 48.1%, 44.0% and 37.5% in day 10, respectively. The extracts of E. prolifera extracted with methanol, acetone and ethyl acetate caused cavities, pieces and pigment reduction in cells, and those with chloroform and petroleum ether caused goffers on cells. The extracts of E. prolifera extracted with all the five solvents decreased athletic ability of the cells, among which those extracted with ethyl acetate, chloroform and petroleum ether decreased cell size of test microalgae. The further investigation found that the methanol extracts significantly decreased contents of chlorophyll, protein and polysaccharide in the cells of those microalgae. The inhibitory effect of chlorophyll, protein and polysaccharide contents of four species of microalgae by the methanol extracts was about 51%. On the basis of the above experiments, dry powder of E. prolifera were extracts with methanol, and extracts were obtained. The methanol extracts were partitioned to petroleum ether phase, ethyl acetate phase, n-butanol phase and distilled water phase by liquid-liquid fractionation, and those with petroleum ether and ethyl acetate significantly inhibited the growth of all test microalgae, and the inhibitory effect of four species of microalgae by those two extracts was above 25% in day 10. Our researches expressed that antialgal substances in E. prolifera extracted with methanol were obtained. And two fractions (petroleum ether phase and ethyl acetate phase) that inhibited the growth of all test microalgae were obtained when the methanol extracts was fractionated by liquid-liquid fractionation.

[Application study of gait analysis on different syndrome types based on musculature zones of the knee osteoarthritis].

OBJECTIVE: To study the gait characteristics of different syndrome types based on musculature zones of the knee osteoarthritis (KOA) and to discuss the application of modern biomechanical technology in objective study on syndrome differentiation which is based on musculature zones. METHODS: Thirty knees out of 19 patients suffering from KOA with the different musculature zone syndrome types were measured the step angle, the heel impulse, the internal and external rotation figures and the percentage of the maximum vertical ground reaction time in the total time of the supporting period by using the Plantar Pressure Measurement System. RESULTS: In the knees with abnormal musculature zones, the figures of step angle and the heel impulse of Foot Shaoyang Meridian and three foot yin meridians were significantly increased than normal (all P<0.05), while those figures of Foot Yangming Meridian were normal; in the knees of Foot Yangming Meridian with abnormal musculature zones, the external rotation mean figure in the midstance was significantly increased than normal (P<0.05); in the knees of Foot Shaoyang Meridian with abnormal musculature zones, the internal rotation mean figure in the propulsion was significantly increased than normal (P<0.05); in the knees of the three foot yin meridians with abnormal musculature zones, the mean external rotation figure in propulsion was significantly increased than normal (P<0.05); in the knees of the Foot Yangming Meridian with abnormal musculature zones, the time of maximum vertical ground reaction was significantly delayed (P<0.05). CONCLUSION: The figures of the gait index in KOA patients with different musculature zone types are different, including the step angle, heel impulse, the internal and external rotation figures, the time of the maximum vertical ground reaction and so on, which could be used as the objective basis of syndrome differentiation based on musculature zones for KOA patients. Therefore, biomechanical technology can be applied in the syndrome differentiation based on musculature zones for KOA patients.

Escin, a natural mixture of triterpene saponins, exhibits antitumor activity against hepatocellular carcinoma.

Escin, a mixture of triterpene saponins extracted from Aesculus wilsonii Rehd., was used to analyze the antitumor effect in hepatocellular carcinoma in vivo and in vitro. At a dose of 2.8 mg/kg, escin had a rather high inhibition ratio (43.5 %) on mice H22 tumor growth in vivo. The results of the SRB cell viability assay showed that escin could induce significant concentration- and time-dependent inhibition of HepG (2) cell viability. Disruption of the G (1)/S phase of cell cycle progression accompanied by the induction of apoptosis were also observed in HepG (2) cells following escin treatment. The results of pulse-field gel electrophoresis and Western blot analysis show the induction of caspase-independent apoptosis by escin. This study provides evidence that escin induces cell cycle checkpoint arrest and caspase-independent cell death in HepG (2) cells, in support of its efficacious potential as a chemopreventive agent.

[Regulative effects of aquaporin 4 expression by rhein in rhubarb to intestinal epithelial cell line LoVo].

OBJECTIVE: To investigate the effects of rhein on regulating aquaporin4 (AQP4) to LoVo cells cultured with RPMI-1640 medium containing rhein. METHODS: LoVo cells were cultured with RPMI-1640 medium containing different concentration rhein for 24 hours and were cultured with RPMI-1640 containing rhein (20 mg/L) for different time. Four groups were assigned as LoVo cells were cultured respectively with RPMI-1640 medium containing different concentration rhein (40, 20, 10 mg/L and control group), while six groups were assigned as LoVo cells were cultured with RPMI-1640 medium containing rhein (20 mg/L) for different time (3, 6, 12, 24, 48 h and control group). The location of AQP4 protein in LoVo cells was definited by immuocytochemistry dying. Western blotting and semiquantitative RT-PCR were adopted to detect the relative expression of AQP4 protein and mRNA. RESULTS: AQP4 was located mainly in plasma membrane of LoVo cells while partly in cytoplasm. The relative expression of AQP4 protein and mRNA decreased with the increasing of rhein concentration; there was no significant difference of the relative expression of AQP4 in 10 mg/L group compared with that in control group, but it decreased significantly in 40 and 20 mg/L groups. The relative expression of AQP4 in 3 and 6 h groups was lower than that in control group but there was no statistical significance, however that in 12, 24, 48 h groups was lower significantly compared with that in control group. CONCLUSION: Rhein can inhibit the genetic transcription and the translation of AQP4 gene in LoVo cells, which demonstrates that the change of AQP4 expression regulated by rhein may be related to the cathartic effect of rhubarb.

[Effect of total anthraquinone in rheum on aquaporin 2 expression in rat distal colon].

OBJECTIVE: To investigate effect of total anthraquinone in rheum on aquaporin 2 expression in rat distal colon. METHOD: SD rats were randomly divided into control group, low dose group, middle dose group and high dose group. Gavaged to control group, and treated group were administered saline and total anthraquinone in rheum with dosage of 0.14, 2.5, 4.5 g x kg(-1) x d(-1), respectively. All rats were put sacrificed after 5 days and stool in full length colon was gently collected to detect water content stool. Distal colon was removed to detect AQP2 expression with immunohistochemistry, western blot and RT-PCR. RESULT: No diarrhea was found in low dose group and control group, there were not significant difference water content of stool and AQP2 expression between low dose group and control group. However, soft feces and loose stools occurred in diarrheic dose group, loose stools and watery stool appeared in high dose group. Stool water content increased in diarrheic dose group and High dose group, expression of AQP2 decreased evidently in these two groups (P < 0.01). CONCLUSION: Total anthraquinone in rheum can reduce the transcription and translation of AQP2 in rats' distal colon, increase fecal water content, which probably is one of the mechanisms of diarrhea caused by total anthraquinone in rheum.

[Effect of electroacupuncture on plasma estrin and bone mineral density in ovariectomized rats].

OBJECTIVE: To observe the effect of electroacupuncture (EA) on plasma estrin (E) 2 and bone mineral density (BMD) in ovariectomized (OVX) rats for studying its underlying mechanism in treating osteoporosis. METHODS: Thirty-two female SD rats were randomized into normal control, model, EA, and medication groups with 8 rats in each group. Postmenopausal osteoporosis model was established by removing the uterus under anesthesia (2% Phenobarbital, 40 mg/kg). In EA group, bilateral "Zusanli" (ST 32) and "Sanyinjiao" (SP 6) were punctured and stimulated electrically for 20 minutes with 1-3 Hz in frequency, 1 ms in duration of waves, and 0.7-1.0 mA in strength, once daily and 8 weeks altogether. Rats of medication group were drenched with 5% Nilestriol, 5 mL/week and for 8 weeks. At the end of experiments, blood samples were collected after removing the rat eyeball, and the left femoral bone tissue was taken. Serum E2 was assayed by using enzyme linked immunosorbent assay (ELISA) and BMD was measured by using double functional X-ray digital bone density meter. RESULTS: Compared with normal group, the body weight of model group was significantly bigger (P < 0.05), and that of model group was also significantly bigger than that of EA and medication groups (P < 0.11). No significant differences were found among the 4 groups before experiments and among normal control and EA groups after treatment (P > 0.05). In comparison with normal group, BMD and serum E2 of model group decreased significantly (P < 0. 01), while compared with model group, BMD and E2 of EA and medication groups increased significantly (P < 0.01, < 0.05). No significant differences were found among normal, EA and medication groups (P > 0.05). CONCLUSION: Both EA and medication can increase BMD and serum E2 in OVX rats, which may be one of the mechanisms of acupuncture in treating osteoporosis.