Salidroside Promotes Sensitization to Doxorubicin in Human Cancer Cells by Affecting the PI3K/Akt/HIF Signal Pathway and Inhibiting the Expression of Tumor-Resistance-Related Proteins.

Salidroside (Sal), the major active constituent of Rhodiola rosea L., is considered as a potential pro-drug with various activities; however, its role in tumor therapy is not clear. Here, we demonstrated in vitro and in vivo that Sal enhanced the inhibitory activity of doxorubicin (DOX) in drug-resistant cancer cell lines. Our results showed that combination drug treatment (Sal and DOX) significantly decreased cell proliferation, migration, and motility. Besides biological validation, a luciferase-labeled animal tumor xenograft model and bioluminescence imaging (BLI) were applied for assessing the tumor progression. Sal combined with DOX inhibited the growth of HeLa-ADR-luc cells in vivo and downregulated the DOX-induced high expression of MDR1. Also, Sal downregulated the Bcl-2, MMP-2, MMP-9, PI3K, and AKT and upregulated BAX proteins. Sal demonstrated high safety and cardiac protection activity. We discovered that Sal enhances DOX sensitivity through the regulation of PI3K/Akt/HIF-1α and DOX-induced resistance pathways. Our results suggest that Sal could be a novel chemosensitization agent for the treatment of multi-drug-resistance tumors.

Blumeaenes A-J, sesquiterpenoid esters from Blumea balsamifera with NO inhibitory activity.

Chemical examination of the ethanol extract of the aerial parts of Blumea balsamifera led to the isolation of ten new sesquiterpenoid esters, blumeaenes A-J (1- 10), with 13 known flavonoids. Their structures were determined mainly by use of 1D and 2D NMR spectroscopic techniques. All sesquiterpenoid esters were tested for their inhibitory activity against LPS-induced NO production in RAW264.7 macrophages. Compounds 1, 4 and 5 showed slight inhibitory effect on the production of NO with IC(50) values of 40.06, 46.35 and 57.80 microg/mL, respectively.

Japonicones E-L, dimeric sesquiterpene lactones from Inula japonica Thunb.

Eight new dimeric sesquiterpene lactones (japonicones E-L, 1- 8), including a novel sesquiterpene dimer bearing a rare hydroperoxide group (japonicone E, 1), were isolated from the aerial part of Inula japonica Thunb. Their structures were determined mainly by the use of 1D and 2D NMR spectroscopic techniques including HSQC, (1)H-(1)H COSY, HMBC, and NOESY. All the isolates were tested for inhibitory effects against LPS-induced NO production in RAW264.7 macrophages. Among the compounds tested, japonicone F (2) showed the strongest activity with the IC(50) value of 4.1 microg/mL.

Japonicones A-D, bioactive dimeric sesquiterpenes from Inula japonica Thunb.

Four new dimeric sesquiterpene lactones japonicones A-D (1-4), comprised by eudesmane and guaiane sesquiterpenes, were isolated from the aerial part of Inula japonica Thunb. The structures and stereochemistry of 1-4 were elucidated by use of 2D NMR spectroscopic techniques, X-ray crystallography and modified Mosher method. Japonicone A (1) showed the most potent cytotoxicities against four tumor cell lines, A549, LOVO, CEM and MDA-MB-435.

A new triterpenoid from Brucea javanica.

A new triterpenoid, bruceajavanin C (1), together with bruceosides A and B (2 and 3), bruceines D and E (4 and 5), yadanziosides A and G (6 and 7), (20R)-O-(3)-alpha-L-arabinopyranosylpregn-5-ene-3beta,20-diol (8), and alpha-D-glucopyranoside, (3beta, 20R)-3-hydroxypregn-5-en-20-yl (9) were isolated from the aerial parts of Brucea javanica. The structure of 1 was elucidated on the basis of 2D-NMR spectroscopic analysis. In addition, compounds 1, 3, 4, 5, and 6 exhibited mild inhibitory effect on NO production in LPS-stimulated RAW264.7 cells.

[Monitoring the synthesis of N,N-diphenyl urea by IR-online].

IR-online detector, React IRTM 4000, was used to monitor the synthesis of N,N-diphenyl urea from urea and aniline. The concentration changes of the reaction components were obtained according to the characteristic IR absorbance changes of the reactants (urea and aniline) at 1 420 and 1 270 cm(-1) respectively, the intermediate product (monophenylurea) at 1 339 cm(-1) as well as the product (N,N-diphenyl urea) at 1 312 cm(-1). In the present study, the result showed that the synthesis of N,N-Diphenyl urea was carried out by two steps. Firstly, the intermediate product monophenylurea was synthesized from urea and aniline, and then N, N-diphenyl urea was obtained by the reaction of monophenylurea with aniline. It was also shown that the IR-online detector was a fast, simple and exact technique to optimize the reaction time and so the reaction course could be easily controlled. The result of IR-online spectrometry was approved by offline reference high performance liquid chromatography (HPLC) method. As a result, the IR-online spectrometry was superior to HPLC method since it was a nondestructive method without the need to sample the reaction medium.

[Value of the antigen with molecular mass of 32 000 in immunodiagnosis of Angiostrongylus cantonensis].

OBJECTIVE: To evaluate the specificity and sensitivity of immunodiagnosis with the antigen with molecular mass of 32 000 of Angiostrongyliasis cantonensis(AC32). METHODS: The major antigenic protein AC32 was purified from the antigens of A.cantonensis adult worm by electroelution from SDS-polyacrylamide gels. AC32-enzyme linked immunosorbent assay (ELISA) and adult worm antigen (AWA)-ELISA were used to detect the specific IgG in the sera of normal rats (n=5) and rats with angiostrongyliasis (n=61), sera of healthy individuals (n=50) and patients (n=50) with angiostrongyliasis, schistosomiasis and other parasitosis. RESULTS: The positivity rate in AC32-ELISA of the sera from rats with angiostrongyliasis was 100%; without false positive results or cross reaction between AC32 and the sera of the normal control and patients infected with parasites other than A.cantonensis. CONCLUSION: AC32 of A.cantonensis is a valuable candidate antigen for immunodiagnosis of angiostrongyliasis with high sensitivity and specificity.

[Expression of a laccase gene from Pleurotus ostreatus in Pichia pastoris and characterization of the recombinant enzyme].

A Pleuotus ostreatus laccase gene was cloned by RT-PCR and designated as lccPol. Its sequence was submitted to GenBank with the accession number AY450404 obtained. The open reading frame was transformed into three Pichia pastoris strains GS115, KM71 and SMD1168, respectively, under control of the AOX1 promoter by using the vector pHBM906. LCCPo1 can be expressed by all three P. pastoris recombinant strains. Three different strategies for shake-flask cultures were compared: (1) (25 degrees C, 1.0% methanol), (2) (20 degrees C, 1.0% methanol), (3) (20 degrees C, 0.5% methanol). The laccase activity could be improved by increasing the methanol concentration befittingly. The results showed that the cultivation temperature had a marked effect on the production of active heterologous laccase. 2 - 6 folds higher laccase activities were obtained when the cultivation temperature was kept at 20 degrees C instead of 25 degrees C. The highest activities, 3.19U/mL [GS115 (pHBM565)], 2.56U/mL [KM71 (pHBM565)], and 2.49U/mL [SMD1168 (pHBM565)], were gotten when the induction were performed at 20 degrees C with 1.0% (V/V) methanol supplied. The temperature and pH optimum for the recombinant laccase produced by three strains were 60 degrees C and pH4.2, respectively.

[Diagnostic value of serum procalcitonin and C-reaction protein in acute exacerbation of chronic bronchitis].

OBJECTIVE: To evaluate the diagnostic value of serum procalcitonin (PCT) and C-reactive protein (CRP) in patients with acute exacerbation of chronic bronchitis. METHODS: The PCT and CRP levels were detected in 56 patients with acute exacerbation of chronic bronchitis and 58 healthy control subjects. RESULTS: The serum PCT level and positivity rate were significantly higher in the case group than in the control group (P<0.05); the serum CRP level was also significantly higher in the case group (P<0.05), but the positivity rate of CRP was similar between the two groups (P>0.05). The sensitivity, specificity, positive predictive value, negative predictive value and diagnostic accuracy rate of PCT were higher than those of CRP. CONCLUSION: Serum PCT is more sensitive than CRP in the diagnoses of acute exacerbation of chronic bronchitis, and can be used as a specific indicator for monitoring the condition of the patients.

Qualitative and quantitative determination of the major coumarins in Zushima by high performance liquid chromatography with diode array detector and mass spectrometry.

A method, high-performance liquid chromatography coupled with diode array and electrospray tandem mass spectrometry (HPLC-DAD-ESI-MS), was developed to qualitatively identify and quantitatively determine the 10 major active coumarins of Zushima. The analysis was performed by using a ZORBAX SB-C18 analytical column (250 mm x 4.6 mm ID, 5 microm) at gradient elution of 0.5% aqueous formic acid and acetonitrile with diode array detection (325 nm). The method was validated for linearity, precision, accuracy, limit of detection and quantification. The proposed method was successfully applied for the qualitative and quantitative analysis of 10 coumarins in five different species of Zushima which had great variation on the contents of investigated coumarins.

Two new coumarins from Edgeworthia chrysantha.

Two new coumarins, 8-[3-(2,4-benzenediol)-propionic acid methyl ester]-coumarin-7-beta-D-glucoside (1) and 7-hydroxyl-odesmethoxyrutarensin (2), were isolated from the stems and barks of Edgeworthia chrysantha. Their structures were elucidated on the basis of spectroscopic data interpretation.

Validation of VKORC1 and CYP2C9 genotypes on interindividual warfarin maintenance dose: a prospective study in Chinese patients.

OBJECTIVES: To develop a warfarin-dosing algorithm that could be combined with pharmacogenomic and demographic factors, and to evaluate its effectiveness in a randomized prospective controlled clinical trial. METHODS: A pharmacogenetics-based dosing model was derived using retrospective data from 266 Chinese patients and multiple linear regression analysis. To prospectively validate this model, 156 patients with an operation of heart valve replacement were enrolled and randomly assigned to the group of pharmacogenetics-guided or traditional dosing for warfarin therapy. All patients were followed up for 50 days after initiation of warfarin therapy. The log-rank test was compared with the time-to-event (Kaplan-Meier) curves. Cox proportional hazards-regression model was used to assess the hazard ratio of the time to reach stable dose. RESULTS: The linear regression model derived from the pharmacogenomic model correlated with 54.1% of warfarin dosing variance. The final multiple linear regression model included age, body surface area, VKORC1, and CYP2C9 genotype. The study showed that the hazard ratio for the time to reach stable dose was 1.932 for the traditional dosing group versus the model-based group and a close and highly significant relationship was observed to exist between the predicted and the actual warfarin dose (R=0.454). CONCLUSION: A pharmacogenetics-based dosing algorithm has been developed for improvement in the time to reach the stable dosing of warfarin. This model may be useful in helping the clinicians to prescribe warfarin with greater safety and efficiency.

Anti-inflammatory and analgesic activities of Edgeworthia chrysantha and its effective chemical constituents.

The barks and roots of Edgeworthia chrysantha LINDL., which have been used as the folk medicine "Zhu shima" in southern China due to their detumescence and acesodyne effects, were investigated for their anti-inflammatory and analgesic activities using a xylene-induced ear edema assay in mice and Freund's complete adjuvant-induced paw edema as inflammation models, and the acetic acid-induced writhing test as an analgesic model. Fractions effective in terms of anti-inflammatory and analgesic activities were obtained from E. chrysantha. The chloroform-soluble fraction (CHF) showed significant anti-inflammatory (p<0.01-0.001) and analgesic (p<0.01) effects. On further purification by silica gel, three major coumarins, edgeworin (EdN), edgeworosides A and C (EdeA and EdeC), were isolated from the chloroform fraction and both anti-inflammatory and analgesic activities were evaluated. EdN and EdeA had anti-inflammatory (p<0.05-0.01) and analgesic (p<0.001) effects, while EdeC only showed an analgesic effect. The results of this study thus demonstrated that the coumarins EdN, EdeA and EdeC in this plant may be active constituents that contribute to the anti-inflammatory and analgesic effects.

Ainsliatrimers A and B, the first two guaianolide trimers from Ainsliaea fulvioides.

Ainsliatrimers A (1) and B (2), the first two guaianolide-type sesquiterpene lactone trimers, together with one new structurally related sesquiterpene dimer ainsliadimer B (3), were isolated from the aerial part of Ainsliaea fulvioides. Their structures were elucidated by spectroscopic techniques, including X-ray crystal diffraction. Both 1 and 2 showed potent cytotoxicites against LOVO and CEM cell lines.