Feasibility exploration of discriminating red yeast rice for different applications by secondary metabolite fingerprints / 药学学报

Though red yeast rice (RYR) has been used as medicine for centuries, few study has been reported about its biological activities related to traditional medicinal application and marketed RYR showed poor consistency in quality. In this study, with comprehensive investigation of their production processes and field acquisition samples including those from genuine producing area, an ultra performance liquid chromatographic (UPLC) method was firstly established to discriminate RYR for different applications based on their secondary metabolites fingerprint. It was performed on a CAPCELL CORE AQ column (100 mm×4.6 mm, 2.7 μm), with PDA (range: 200-650 nm, extracted: 237 nm) and ELSD detection. The mobile phase used was water (A) and acetonitrile (B) both containing 0.1% formic acid at gradient elution (0-15 min, 50% B→85% B (linear); 15-16 min, 85 % B→50% B (linear) and maintained until 21 min), with a flow rate of 0.5 mL∙min-1. The method established was fully validated in agreement with guidelines of Chinese Pharmacopeia. Common metabolites were found in RYR for same application and the fingerprints of RYR for food coloring or brewing from various manufacturers had similarities above 0.90. Meanwhile, significant differences were observed among the fingerprints for various applications and discrimination could be achieved by principal component analysis (PCA). Lovastatin was absence in RYRs for food coloring or brewing, and the fingerprint of traditional medicinal RYR was similar to that of RYR for brewing. However, standardization was required for RYR containing lovastatin because of their significant differences from various manufacturers in fingerprints and lovastatin content. The results demonstrated the feasibility to discriminate RYR for different applications by the secondary metabolites fingerprint method established in this study, which provides a scientific basis to investigate the relationship between biological activities of medicinal RYR and their corresponding secondary metabolites, and further aid their quality standardization and improvement.

Study on Stability of Mixture Reference Solution of Strychni Alkaloids / 广州中医药大学学报

Objective Brucine and strychnine monomer reference substance as extremely toxic substance had potential threat during transportation and utilization. In this study we investigated the homogeneity, stability and assignment accuracy of the mixture reference solution of strychnine and brucine, so as to provide reference for the quality control of extremely toxic chemical reference substances for Chinese medicine. Methods Following the assay in Chinese Pharmacopoeia volume I (2015), we prepared the mixture reference solution of brucine and strychnine, and investigated the solvents and the concentration of mixutre reference solution. The stability test lasted for 12 months. F-test was used for heterogeneity assay. Three researchers were involved for collaboration. Results Methanol and chloroform solution were selected as the solvents for the stability test. Results showed the difference was not statistically significant among various mixture solutions. The results of value assignment were 0.14 mg/mL for strychnine (sR = 0.5％)and 0.10 mg/mL for brucine (sR = 1.0％). The stability of mixture solution were better under the conditions of methanol solution at 4 ℃ or -20 ℃. Conclusion The results provide a possible way to develop the mixture solution in place of the monomer reference, and the mixture reference solution is expected for the quality control in the slices of Semen Strychni and its compound preparations.

Expression of c-IAPl protein and mRNA in gastric adenocarcinoma and its clinical significance / 临床与实验病理学杂志

Purpose To investigate the protein and mRNA expression of c-IAPl in gastric adenocarcinoma tissue and its clinical significance. Methods Immunohistochemistry technique, Western blot and realtime fluorescent quantitative PCR(qRT-PCR) were used to detect the protein and mRNA expression of c-IAPl in 50 cases of gastric adenocarcinoma tissues and40 cases of adjacent normal gastric mucosa tissues to analyze its role in the development and progression of gastric adenocarcinoma, the relationship between the protein and mRNA expression of c-IAPl and the clinicopathological features. Results The relative expression level of c-IAPl protein in gastric adenocarcinoma tissues was significantly higher than that in adjacent normal gastric mucosa tissues, the difference was statistically significant (P＜ 0.05 ). The mRNA expression of c-IAPl in gastric adenocarcinoma was significantly higher than normal gastric mucosa tissues, the difference was statistically significant (P＜0.05). The protein and mRNA expression of c-IAPl were correlated with the degree of differentiation of gastric adenocarcinoma tissues, TNM clinical stage, lymph node metastasis and infiltration depth, the difference was statistically significant (P＜0.05 ), while there was no correlation with gender and age, the difference was not statistically significant (P＞ 0.05). Conclusion The high protein and mRNA expression of c-IAPl in gastric adenocarcinoma tissues inhibit the apoptosis of gastric adenocarcinoma cells, which contribute to the development and progression of gastric carcinoma and it may provide a new theoretical basis for the clinical targeted therapy of gastric adenocarcinoma.

Impurity profiling of macrolide antibiotics by liquid chromatography-mass spectrometry / 药学学报

Macrolide antibiotics are broad-spectrum, with activity against a range of Gram-positive, Gram-negative organisms and some anaerobes. The components of macrolide antibiotics are generally complicated. Therefore, it is very important to establish impurity profiles of these antibiotics to ensure their safety and process control. Compared with classical methods, the liquid chromatography-mass spectrometry method is particularly advantageous to characterize minor components at trace levels in terms of sensitivity, efficiency and selectivity, thus more and more widely used in establishments of impurity profiles. In this study, the general approaches to characterize minor components in complex pharmaceutical matrix, fragmentation pathways of 14- and 16-membered macrolide antibiotics and the establishment of the impurity profile of acetylspiramycin were given to provide valuable enlightenments to establish the impurity profiles of pharmaceutical products.

Stability of rh-endostatin and fabrication/release test of rh-endostatin loaded PLGA microspheres / 第二军医大学学报

Objective: To prepare rh-endostatin loaded poly (lactic-co-gtycolic acid) (PLGA) microspheres and to evaluate their release behavior in vitro. Methods: Rh-endostatin PLGA microspheres were prepared by W/O/W process. The content and in vitro cumulative release was determined by a HPLC method. Results: The prepared microspheres were well-shaped, with a mean diameter of 122.7 μm. The drug loading and encapsulation efficiency were 1.28% and 38.65%, respectively. The rh-Endo solution (250 μg/ml) showed good stability after placed at 4°C and 25°C for 108 h. The cumulative in vitro release was up to 67.37% in 28 days. Conclusion: The rh-Endo can be encapsulated in microspheres to yield sustained release using biodegradable polymers PLGA as the carrier material.

In vivo anti-tumor effect of Oxytropis kansuensis alkaloid fraction and its influence on immune function / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To study the in vivo anti-tumor effect of Oxytropis kansuensis alkaloid fraction (OKAF) in live cancer cell line H22 and its influence on immune function in KM mice.</p><p><b>METHODS</b>Sixty mice with transplanted mouse originated liver cancer cell line H22 were divided randomly into five groups, the three OKAF groups were administrated with OKAF in low (8 mg/kg), moderate (16 mg/kg) and high (32 mg/kg) dosage, the negative control group was administrated with normal saline 20 mL/kg and the positive control with 5-FU 15 mg/kg, respectively, once a day for 10 successive days. Then the average tumor weight (TW), tumor inhibition rate (TIR), spleen index (SI), and thymus index (TI) in them were calculated, and the spleen cell proliferation rate (SCPR) was measured by MTT assay.</p><p><b>RESULTS</b>TIR of low, moderate and high dosage of OKAF was 23%, 29% and 43% respectively. SI and TI in all the three OKAF groups were higher than those in the negative control group, and the two indexes in the moderate and ligh dosage OKAF groups were higher than those in the positive control group (P < 0.05). SCPR in OKAF groups at 24 h, 48 h and 72 h were higher than those in both two control groups at the same time.</p><p><b>CONCLUSION</b>OKAF shows inhibitory action on liver cancer cell line H22 tumor in KM mice, and it could also enhance the immune function of mice.</p>

Analysis of the response factors of different aminoglycoside antibiotics detected by evaporative light-scattering detector / 药学学报

<p><b>AIM</b>To analyze if the response factors of different aminoglycoside antibiotics detected by evaporative light-scattering detector (ELSD) are the same. If they are, then ELSD can be applied to the quality analysis of this class of antibiotics.</p><p><b>METHODS</b>The response factors of five different aminoglycosides (amikacin, sisomicin, netilmicin, etimicin and vertilmicin) detected by ELSD were determined by using a Diamonsil C18 column (150 mm x 4.6 mm, 5 microns) as analytical column and 0.2 mol.L-1 trifluoroacetic acid-methanol (94:6) as mobile phase at a flow rate of 0.6 mL.min-1, the temperature of the drift tube was set at 110 degrees C, and the flow of carrier gas at 2.80 L.min-1. Detector responses (A) and the amount of injection of each substance (m) were fitted to the logarithmic regression: logA = blogm + loga.</p><p><b>RESULTS</b>The linear regression equation obtained were amikacin: Y = 1.46X + 5.07, gamma = 0.9997; sisomicin: Y = 1.51X + 5.03, gamma = 0.9997; netilmicin: Y = 1.52X + 4.88, gamma = 1.000; etimicin: Y = 1.46X + 4.85, gamma = 0.9999; vertilmicin: Y = 1.41X + 4.90, gamma = 0.9998. The differences between them were negligible.</p><p><b>CONCLUSION</b>Different aminoglycosides can give the same responses with ELSD detection. So, the HPLC-ELSD methods can be applied to the analysis of impurities, the control of the ratio of multi-components drug and the determination of new substances by using another substance as reference, etc.</p>