Effects of different drying conditions on protein in Cordyceps / 中国中药杂志

In this study,the protein in different Cordyceps samples,which include fresh sample( S1),22 ℃ drying sample( S2),37 ℃ drying sample( S3) and 60 ℃ drying sample( S4),were analyzed by sodium dodecylsupinate-polyacrylamide gel electrophoresis( SDS-PAGE) and two-dimensional electrophoresis( 2-DE). The total protein contents in Cordyceps samples were from 1. 655-4. 493 mg·g~(-1) and the protein contents in fresh sample was the highest. The results of SDS-PAGE showed that the mainly ranges of protein molecular weight of Cordyces samples were 10-100 kDa and the numbers of protein bands were 28 to 41,the fresh sample had the maximum number of protein bands. The 2-DE profiles were analyzed by PDQuest software. The resulted indicated that 488-876 protein spots were detected in different Cordyceps samples and the isoelectric point( pI) was distributed between 4. 5 and 6. 5,the protein molecular weight was distributed in 10-20 kDa and 25-100 kDa,the fresh sample had the maximum number of protein spots. Therefore,the drying process could decrease contents and species of protein in Cordyceps,and the different drying conditions had different effects on protein. These results provide a reference for improving the drying process of Cordyceps.

Synthesis and Release of Dapsone-Alginate (DS-Alg) Conjugate / 中国药学杂志

OBJECTIVE: To synthesize and characterize dapsone-alginate acid (DS-ALG) conjugate. METHODS: Alginate (Alg) was selected as the drug carrier and valine (Val) as the linking arm to synthesize DS-Alg, which could be applied to topical administration. And the synthetic condition of DS-Alg was optimized by changing the amount of 1-(3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride (EDC) and N-hydroxysuccinimide (NHS), while the pH of solvent was changed in the range of 4.0 to 6.0. The structures of the products were characterized by 1H-NMR, MS and FT-IR. Meanwhile, the drug release in vitro of DS-Alg was investigated in the mixture of pH 7.4, 0.05 mol•L -1 PBS and ethanol by diffusion cells. The concentration of DS or valine-dapsone in the release medium was detected by HPLC. Taking rats with local scald as model, the drug release in vivo was measured by coating the trauma with DS-Alg conjugate cream and monitoring the drug concentration in blood. RESULTS: The optimum synthetic conditions of DS-Alg were as follows: 0.277 g valine-dapsone, 0.400 g sodium alginate, 7 eq EDC, 3 eq NHS, pH of the solvent of 5.5. And during 72 h, there was no DS detected in the release medium or rat plasma. The AUC0→72 h of DS-Alg was 0. It suggested that DS immobilized by Alg with covalent bond was too stable to be released from Alg in vitro and in vivo. The DS-Alg conjugate could effectively prevent DS from entering the systemic circulation. CONCLUSION: DS-Alg conjugate is successfully synthesized. The conjugate is stable that DS cannot be released from the conjugate to the bloodstream, which can efficiently decrease the side effects of DS and has the potential for topical administration.

A lipophilic prodrug of Danshensu: preparation, characterization, and in vitro and in vivo evaluation / 中国天然药物

Danshensu [3-(3, 4-dihydroxyphenyl) lactic acid, DSS], one of the significant cardioprotective components, is extracted from the root of Salvia miltiorrhiza. In the present study, an ester prodrug of Danshensu (DSS), palmitoyl Danshensu (PDSS), was synthesized with the aim to improve its oral bioavailability and prolong its half-life. The in vitro experiments were carried out to evaluate the physicochemical properties and stability of PDSS. Although the solubility of PDSS in water was only 0.055 mg·mL, its solubility in FaSSIF and FeSSIF reached 4.68 and 9.08 mg·mL, respectively. Octanol-water partition coefficient (log P) was increased from -2.48 of DSS to 1.90 of PDSS. PDSS was relatively stable in the aqueous solution in pH range from 5.6 to 7.4. Furthermore, the pharmacokinetics in rats was evaluated after oral administration of PDSS and DSS. AUC and t of PDSS were enhanced up to 9.8-fold and 2.2-fold, respectively, compared to that of DSS. C was 1.67 ± 0.11 μg·mL for PDSS and 0.81 ± 0.06 μg·mL for DSS. Thus, these results demonstrated that PDSS had much higher oral bioavailability and longer circulation time than its parent drug.

Application of computer color matching technology in color simulation of Ruyi Jinhuang Powder placebo / 中草药

Objective To study the color of Ruyi Jinhuang Powder (RJP) placebo by using computer color matching technology and achieve fast preparation of RJP placebo. This work provided a new method to simulate the color of Chinese medicine compound placebo. Methods Using the RJP placebo as an example, computer color matching technology was used to establish a mathematical model to correlate the placebo color parameter L, a*, b* value and the colorant concentration. For a measured medicine compound color, the placebo colorant concentration can then be calculated from the model through data fitting and Newton iterative method. The model accuracy was validated using the color comprehensive evaluation index (ΔE). Results The color parameter L, a*, b* of RJP placebo was 68.302 5, 4.079 5, and 34.484 0. The mass fraction of lemon yellow, amaranth, and blue was 0.837 3%, 0.045 8%, and 0.008 5%. The ΔE value between RJP and its placebo was 2.750 0 ± 0.353 6, and there was no obvious visual difference between the medicine compound and its placebo. Conclusion Computer color matching technology can be used to simulate the color of RJP placebo, and can be widely used in the preparation of Chinese medicine placebo.

Evaluation for extraction process of Shenqi Tongmai Yizhi particles based on antioxidant capacity in vitro and its spectrum-effect relation / 中国中药杂志

Based on DPPH method, the antioxidant activities of Shenqi Tongmai Yizhi particles with different extraction processes were compared. The contribution to the anti-oxidant capacity in vitro was explored by means of grey relational analysis on different chemical compositions in the fingerprint. The results showed that the IC₅₀ concentration values of water extract, water extract from alcohol precipitation, alcohol extract, and alcohol and water extract were 0.801 4, 0.859 1, 0.796 1, 0.918 0 g•L⁻¹; and the alcohol extract is the best method to extract antioxidative components, with the highest antioxidant activity and lowest IC₅₀. When the mass concentration of the herbs reached a certain degree, its free radical clearance rate was similar to that of vitamin C control group. The order of different chemical contributions of constituents to the antioxidant activity in the fingerprint was 4>3>33>53>9>10>11>34>15>59>8>61>52>20>42>18>29. The preliminary exploration for the spectrum efficiency relations provides reference for studying traditional Chinese medicine compound processing method and the pharmacodyamic material basis.

Regeneration of Carthamus tinctorius from Jimsar / 中草药·英文版

Objective: To investigate the factors significantly affecting the shoot buds regeneration of Carthamus tinctorius from Jimsar (CTJ). Methods: Through tissue culture experiments, the influences of culture temperature, illuminance, relative humidity, explants type, seedlings age, and plant growth regulators added in culture media on in vitro regeneration of CTJ were investigated. Results: The culture temperature was set at 24 °C in daytime and 16 °C at night, illuminance at 9000 lx, and 60% relative humidity were suitable for CTJ regeneration. Cotyledons excised from 6-8 d old seedlings were more responsive than even-aged euphyll, hypocotyl, and root explants because of inducing adventitious buds. The highest percentage of regenerated shoots (79.1%) with about five adventitious buds per responding explant was obtained from MS basal medium containing 12.0 mg/L TDZ, 2.5 mg/L IBA, and 1.5 mg/L 2-ip (No. 14 medium). Regenerated shoot buds (80%) could elongate successfully after 1-2 weeks transferred to shoot elongation medium. Conclusion: Suitable factors of shoot buds regeneration for CTJ were determined. It may also make a useful reference for regeneration researches of other C. tinctorius varieties in China. © 2014 Tianjin Press of Chinese Herbal Medicines.

Effects of carbon components of fine particulate matter (PM2.5) on atherogenic index of plasma / 中华预防医学杂志

<p><b>OBJECTIVE</b>To evaluate associations between carbon constituents of fine particulate matter (PM2.5) and atherogenic index of plasma (AIP).</p><p><b>METHODS</b>We collected subjects from two communities by a system sampling, and 112 people aged over 60 years old without cardiovascular disease were recruited. The levels of cholesterol (TC), triglycerides (TG), high-density lipoprotein cholesterol (HDL-C), low density lipoprotein cholesterol (LDL-C) of objects, and personal exposure to PM2.5 were measured on December, 2011. Total carbon (TC), organic carbon (OC) and elemental carbon (EC) of PM2.5 were detected and AIP was calculated according to its definition.</p><p><b>RESULTS</b>The value of AIP among the 112 subjects was 0.05 ± 0.26. Personal exposure concentration of PM2.5 and its carbon components (TC,OC and EC) were (164.75 ± 110.67), (53.86 ± 29.65), (44.93 ± 26.37) and (9.49 ± 5.75) µg/m(3), respectively. The Pearson analysis showed the linear relationship between TC,OC,EC and AIP, all significant positive correlations. The correlation coefficients were TC (r = 0.307, P < 0.05),OC (r = 0.287, P < 0.05) and EC (r = 0.252, P < 0.05), respectively. The multiple logistic regression analysis showed that when the AIP risk categories were selected as dependent variable and low risk group as reference group, the regression coefficient of TC,OC and EC was separately 1.03 (95%CI:1.01-1.05), 1.03 (95%CI:1.01-1.05), 1.12 (95%CI:1.02-1.22) in the high risk group; while there was no statistical significance of the regression coefficient and OR in the middle risk group.</p><p><b>CONCLUSION</b>There was stable associations between the carbon constituents (TC,OC and EC) of fine Particulate Matter (PM2.5) and AIP. The findings suggested that carbon components of PM2.5 should be considered as risk factors of atherogenic.</p>

Determination of ellagic acid, flavonoids and goshonoside-F5 in Rubi Fructus by HPLC / 中国中药杂志

High-performance liquid chromatographic coupled with variable wavelength detection (HPLC-VWD) has been developed for simultaneous determination of 5 analytes including ellagic acid, quercetin, kaempferol-3-O-beta-D-rutinoside, tiliroside and kaempferol, and high-performance liquid chromatographic with an evaporative light scattering detector (HPLC-ELSD) has been established to determine goshonoside-F5 in extract of Rubi Fructus. Chromatographic separations were carried out on an Agilent ZORBAX SB-C18 column (4.6 mm x 250 mm, 5.0 microm). All calibration curves of reference standards revealed good linearity (R2 > 0.999 5) within the concentration ranges tested. The method limits of detection ranged 0.297-90.144 ng and the method limits ofquantitation ranged 0.990-300.480 ng, respectively. Recoveries of 6 analytes were from 97.11% to 101.7%, with RSD less than 2.1%. The result shows that amounts of the 6 analytes in the samples from 16 localities were found to be different. The higher latitude of growing environment, the more ellagic acid in herb. The content of total flavonoids in sample from east localities were higher than that in middle and west localities, and the content of goshonoside-F5 in Bozhou, Anhui province was higher than others. This method was found to be simple, accurate, sensitive with good repeatability. Those results might serve as a sound foundation for further study, quality control and application of Rubi Fructus.

Chemical constituents in Akebiae Fructus based on UFLC-Q-TOF/MS / 中草药

Objective: To identify the chemical constituents in Akebiae Fructus by UFLC-Q-TOF/MS method. Methods: The separation was performed on an Acquity UPLC BEH C18 Column (100 mm × 2.1 mm, 1.7 μm), with a mobile phase using 0.1% formic acid-acetonitrile and water containing 0.1% formic acid (B) for gradient elution. The flow rate was 0.3 mL/min, the temperature of column was 40°C with injection volume of 1 μL. TOF/MS and electrospray ion (ESI) source were applied for the qualitative analysis under the negative ion mode, and the full mass scan range was m/z 100-1500. Results: According to MS principle, twenty-five triterpenoids were identified from the methanol extract of Akebiae Fructus and the chemical structures of other nine unknown compounds were deduced. Conclusion: UFLC-Q-TOF/MS method could identify the main chemical constituents in Akebiae Fructus rapidly and accurately, which lays a foundation for the quality control of Akebiae Fructus.

Correlation between atmospheric polycyclic aromatic hydrocarbons exposure and urinary hydroxyl metabolites of polycyclic aromatic hydrocarbons in elderly population in Tianjin, China / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To identify suitable hydroxyl polycyclic aromatic hydrocarbons (OH-PAHs) for co-evaluation of internal exposure level of PAHs by simultaneous determination of a variety of OH-PAHs in urine.</p><p><b>METHODS</b>The 24-h individual particulate matter and morning urine samples of 112 subjects were collected during June 2011. PAHs carried by individual particulate matter samples and OH-PAHs in urine samples were detected by gas chromatography-mass spectrometry.</p><p><b>RESULTS</b>Seven OH-PAHs were detected in urine samples, among which 1-hydroxy-naphthalene (1-OHNap) concentration was the highest [(20.54 ± 28.94) µmol/mol Cr], while 1-hydroxy-pyrene (1-OHP) concentration was the lowest [(0.73 ± 0.63) µmol/mol Cr]. The concentrations of these seven OH-PAHs decreased in the following order: 1-hydroxy-naphthalene (1-OHNap) > 9-hydroxy-fluorene (9-OHFlu) > 2-hydroxy-naphthalene (2-OHNap) > 3-hydroxy-fluorene (3-OHFlu) > 2-hydroxy-fluorene (2-OHFlu) > 6-hydroxy-chrysene (6-OHChr) > 1-hydroxy-pyrene (1-OHP). The effects of gender and smoking upon the contents of OH-PAHs in urine samples were not significant. There was a good correlation between total hydroxy-naphthalene (ΣOHNap) and 1-OHNap (r = 0.948), and a good correlation was also showed between total hydroxy-fluorene (ΣOHFlu) and 9-OHFlu (r = 0.975). Naphthalene carried by atmospheric particulate matters demonstrated better correlation with 1-OHNap than 2-OHNap, while fluorene carried by atmospheric particulate matters showed better correlation with 9-OHFlu than 3-OHFlu and 2-OHFlu. The correlation coefficients of ΣOHNap, ΣOHFlu and 6-OHChr with 1-OHP were 0.427, 0.543 and 0.655, respectively, and the correlations were not strong.</p><p><b>CONCLUSION</b>It cannot reflect internal exposure level of PAHs to use 1-OHP as the only biomarker, while 1-OHNap and 9-OHFlu can be well predictive of the exposure levels of corresponding total OH-PAHs, suggesting that simultaneous determination of 1-OHNap, 9-OHFlu and 1-OHP can be more accurate and comprehensive in evaluating the internal exposure level of PAHs.</p>

Study of flavanoids extracted from onion on the blood-brain barrier permeation and neuroprotective effects / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To study the flavanoids extracted from onion on the blood-brain barrier (BBB) permeation, and their effects on primary cultured neuron cell proliferation and apoptosis of SD rats using ethanol reflux method.</p><p><b>METHODS</b>The brain microvascular endothelial cells (BMVECs) were first successfully primary cultured. Then rats BMVECs and astrocytes (ACs) were co-cultured to establish the in vitro BBB model. The flavanoids were extracted from onion using ethanol reflux method. The model was verified by transmission electron microscopy (TEM) and trans-epithelial electric resistance (TEER). The flavanoids permeability was tested using high performance liquid chromatography (HPLC). Meanwhile, rat neuron cells were cultured and exposed to H2O2 and flavanoids. Their effects on the cell proliferation and apoptosis were observed using MTT assay. The injury of neuron DNA was analyzed using single-cell gel electrophoresis (SCGE) and immunofluorescent assay.</p><p><b>RESULTS</b>The in vitro BBB model was successfully established by TEM and TEER. Results of HPLC proved flavanoids extracts could effectively permeate the BBB with the permeability of 60.58%. The extractive at 10 - 20 microg/mL showed obvious inhibition on the apoptosis of neuron cells induced by H2O2, and attenuated the injury of neuron DNA.</p><p><b>CONCLUSIONS</b>The flavanoids extracted from onion ethanol reflux method could effectively penetrate the BBB. They also showed obvious inhibition on the H2O2 induced neuron cell apoptosis and DNA injury.</p>

Association of XPC and XPG polymorphisms with the risk of hepatocellular carcinoma / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate whether the polymorphism of DNA repair genes XPC (Ala499Val and Lys939Gln) and XPG (His1104Asp) is associated with the susceptibility to hepatocellular carcinoma (HCC).</p><p><b>METHODS</b>A hospital-based case-control study was conducted in 500 cases with HCC and 507 controls. Genotypes of XPC and XPG were determined by real-time polymerase chain reaction with the TaqMan MGB probe.</p><p><b>RESULTS</b>Compared to the CC genotype, the CT genotype and the TT genotype of XPC Ala499Val were not associated with the susceptibility to HCC (adjusted OR = 1.34, 95% CI: 0.85-2.12; adjusted OR = 1.30, 95% CI: 0.68-2.51, respectively). Compared to the AA genotype, the AC genotype and the CC genotype of Lys939Gln were not associated with the susceptibility to HCC (adjusted OR = 1.20, 95% CI: 0.78-1.85; adjusted OR = 1.81, 95% CI: 0.88-3.73, respectively). Compared to the CC genotype, the CG genotype and the GG genotype of XPG His1104Asp were not associated with the susceptibility to HCC (adjusted OR = 0.85, 95% CI: 0.56-1.27; adjusted OR = 1.12, 95% CI: 0.67-1.87, respectively) However, the stratified analysis revealed that the females with the AC+CC genotype of XPC Lys939Gln had increased risk of HCC compared to those with AA genotype (OR = 2.17, 95% CI: 1.01-4.64).</p><p><b>CONCLUSION</b>Our results suggest that XPC and XPG polymorphisms do not independently affect on the susceptibility to HCC, but the joint effect of C allele of XPC Lys939Gln and female may modify the risk of HCC.</p>