OBJECTIVE To establish a self-contrast HPLC method with correction factor for determination of the related substances in tamsulosin hydrochloride, and validate the limits of impurities based on the prediction of genotoxicity using Nexus 2.6 software (with Derek and Sarah). METHODS ZORBAX SB-C₁₈(4.6 mm×150 mm,3.5 μm)column was used for the determination of correction factors of the eight known impurities in tamsulosin hydrochloride with mobile phase consisting of perchlorate buffer solution-acetonitrile by gradient elution under the detection wavelength of 225 nm, the related substances in tamsulosin hydrochloride were determine by self-contrast HPLC method with the correction factor, and the genotoxicity of the impurities was predicted by using Nexus 2.6 software. RESULTS There was no significant difference (the deviation is within ±0.02%) between the results by relative correction factors and external standards. The validation test showed that the proposed method met the requirements for the intended analytical applications, and the predicted result of impurity I by Nexus 2.6 software was positive (ICH M7 class 3), and all the others were negative (ICH M7 class 5). CONCLUSION The established method is rapid, efficient, accurate and sensitive, and a limit of 0.1% is established for each known impurity according to the predicted genotoxicity. This study provides a basis for revising pharmacopoeia standards.

Morinda officinalis is one of the four traditional southern medicines in China. Modern studies show that its oligosaccharides have good anti-depression activity.After continuous research and development by Chinese scientists, Morinda officinalis oligosaccharide Capsules, which are mainly composed of Morinda officinalis oligosaccharide, have been approved to be marketed as the fifth kinds of anti-depression new drugs in Chinese traditional medicine with independent intellectual property rights.By referring to the domestic and foreign literature on Morinda officinalis oligosaccharide as the research object, this paper comprehensively summarizes and combs the research and development process, anti-depression mechanism, extraction process, detection method and clinical application of Morinda officinalis oligosaccharide, which provides references for the subsequent research on Morinda officinalis oligosaccharides and the research and development of antidepressant drugs discovered from traditional Chinese medicine and natural products.

OBJECTIVE To study the effects of different concentrations of photosynthetic induction factor (PIF), methyl jasmonate (MeJA), ceric ammonium sulfate (ACS) and acetylsalicylic acid (ASA) on the accumulation of fucoxanthin in Phaeodactylum tricornutum, screen the best elicitor combination for fucoxanthin accumulation, and study the photosynthetic physiology and gene expression changes of fucoxanthin accumulation in Phaeodactylum tricornutum under the treatment of the best inducer combination. METHODS A four-factor, four-level orthogonal test was used to screen the best inducer combination to promote the accumulation of fucoxanthin in Phaeodactylum tricornutum, and the chlorophyll fluorescence parameters of Phaeodactylum tricornutum under the treatment of the best elicitor combination were detected by pulse-amplitude-modulation (PAM). The expressions of zds, lcyb, rbcL and psbA were detected by RT-qPCR. RESULTS The results of the orthogonal test showed that MeJA was the most significant factor affecting the accumulation of fucoxanthin in Phaeodactylum tricornutum cells, followed by ASA, and the best elicitor combination to promote the accumulation of fucoxanthin in Phaeodactylum tricornutum was 1 μg·L^-1 PIF+0.2 mg·L^-1 ACS+40 mg·L^-1 ASA. The photosynthetic physiological study showed that the inductor treatment was able to increase the fucoxanthin content by 104% compared with the control group, which was 21.77 mg·g^-1. The results of photosynthetic physiological study showed that the inducer combination significantly increased the rETR_max value (P<0.05) of Phaeodactylum tricornutum. The expression of rbcL gene was the highest, which was 2.0 times higher than that of the control treatment (P<0.01). CONCLUSION The expressions of rETR_max, chla content, lcyb, rbcL and psbA genes are highly significantly correlated with zds gene, and the photosynthetic efficiency can be improved by promoting the expression of photosynthetic genes and the relative electron transfer rate of PSⅡ, and the expression of genes related to the synthesis of lutein can promote the accumulation of fucoxanthin in Phaeodactylum tricornutum.

OBJECTIVE To prepare triptolide lignoceric acid ester liposome and characterize it,investigate its therapeutic effect on pancreatic cancer. METHODS Firstly, triptolide lignoceric acid ester liposome was prepared by thin film dispersion method,the single factor test and Box-Behnken response surface method were used to optimize the formulation process. Secondly, liposome form was observed using transmission electron microscope, the particle size, the polydispersity index and Zeta potential of the liposome was observed using Malvin particle size instrument and the initial stability of the TPL-LA-lip was investigated. Finally, the anti-pancreatic activity of TPL-LA-lip in vivo was evaluated by Panc 02 tumor bearing mouse model. RESULTS Triptolide lignoceric acid ester liposome was prepared successfully, and the optimal process and prescription were determined. The liposome was prepared by thin film water method, phospholipid was selected PC-98T, the ratio of drug to phospholipid was 1∶10, cholesterol to phospholipid ratio was 1∶10, DSPE-mPEG2000 was 0.05%, and the preparation temperature was 55 ℃. The liposome was spherical in appearance, the encapsulation rate (EE%) was (98.30±0.32)%, the loading efficiency rate(LE%) was (8.33±0.24)%, the particle size was (105.60±0.01) nm, the Zeta potential was (-34.54±0.17) mV, and had good stability. In PBS medium containing 30% ethanol, the cumulative release of prodrug liposome was 40.35% at 24 h, and it showed good sustained-release effect. At the end of the pharmacokinetics experiment on day 15, the tumor bodies of mice in control, blank lip, TP solution, minnelide and TPL-LA-lip were (849.45±53.72)(880.45±121.45)(602.09±56.80) (265.67±23.12) (237.67±38.30) mm³, respectively. The change rates of body weight were 18.12%, 21.29%, -3.62%, 13.06% and 19.97%, respectively. Compared with the control and TP groups, the TPL-LA-lip group tumor volume was significantly different (P<0.05). In addition, there was no significant difference in body weight between the TPL-LA lip group and the negative control group, while the body weight and organs indexs of the mice treated with TP solution were significantly reduced, indicating the good biological safety of TPL-LA-lip. CONCLUSION The high lipophilic triptolide lignoceric acid ester prodrug greatly improved the formulation druggability of TP, and the liposome produced had high encapsulation rate and good stability. Prodrug technology combined with liposome carrier delivery of TP can significantly enhance the anti-pancreatic cancer effect of the drug, while reducing toxicity, providing a new idea and experimental basis for the development of triptolide prodrug nano drug delivery system.

OBJECTIVE To establish critical quality attributes analysis method for recombinant human collagen Ⅲ. METHODS The recombinant human collagen Ⅲ molecular weight (MW) distribution was determined by matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF MS) and size exclusion chromatography with multi-angle light scattering (SEC-MALS). Capillary electrophoresis-sodium dodecyl sulfate (CE-SDS) and size exclusion high performance liquid chromatography (SE-HPLC) were used for purity analysis. Ion exchange chromatography and capillary zone electrophoresis (CZE) were used to separate and analyze charge isomers. The “double” enzyme digestion peptide mapping method was used for identification and sequence coverage analysis. RESULTS The recombinant human collagen Ⅲ MW distribution measured by MALDI-TOF was 45.01×10³. The MW of monomer recombinant human collagen Ⅲ determined by SEC-MALS was 45.17×10³ (±0.226%). CE-SDS analysis showed that the purity of recombinant human collagen Ⅲ was 98.77% and that of other ingredients was 1.23%. The SE-HPLC purity was 98.07%, and that of other component was 1.93% (dimer). A total of 14 charge isomers were identified through strong cation exchange, and the acidic peak was 52.08%, the main peak was 26.22%, and the basic peak was 21.70%. Eight charge isomers were identified by CZE, including 52.10% acidic peak, 25.27% main peak, and 22.63% basic peak. Two identification methods for charge isomers have consistent distribution patterns. The “double” enzyme digestion peptide mapping method was used for identification and sequence coverage analysis, achieving 100% coverage of the recombinant human collagen Ⅲ sequence. CONCLUSION A series of critical quality attributes analysis methods for recombinant human collagen Ⅲ have been established. These methods provide a reference basis for the quality control of recombinant human collagen Ⅲ in China.

OBJECTIVE To establish UPLC fingerprint of Angelica dahurica cv. Qibaizhi, identify the compounds of common peaks by UPLC-Q-TOF-MS technology, determine antioxidant activity of each batch and study the relationship between effects of antioxidant and spectrum of Angelica dahurica cv.Qibaizhi. METHODS The fingerprint of 28 batches of Angelica dahurica cv. Qibaizhi was established by the Similarity Evaluation System of TCM Chromatographic Fingerprint (2012 edition), and the common peaks were identified and evaluated for similarity, and the common peaks were analyzed by UPLC-Q-TOF-MS. The free radical scavenging rate of 1, 1-diphenyl-2-trinitrophenylhydrazine (DPPH) was used as antioxidant index to determine its antioxidant activity. Grey correlation degree and partial least squares regression were used to analyze the spectral effect relationship between chemical constituents and antioxidant activities of Angelica dahurica cv. Qibaizhi. RESULTS The fingerprints of 28 batches of Radix Angelica were established, the similarity was 0.910-0.997. By cluster analysis, the different processing methods can be divided into two categories: sulfur fumigation and sulphur-free. When the square Euclidean distance is 10, the sulphur-free Angelica dahurica cv. Qibaizhi can be further divided into two categories, which is consistent with the classification of commodity specifications. A total of 13 common peaks were identified. The common peaks were identified by comparison of standards and UPLC-Q-TOF-MS analysis as quinic acid, dihydrooroselol, oxypeucedanin hydrate, byakangelicin, xanthotoxin, bergapten, isopimpinellin, oxypeucedanin, imperatorin, phellopterin, cnidilin, isoimperatorin and falcarinolone. By scavenging DPPH free radicals, it was found that 28 batches of Angelica dahurica cv. Qibaizhi had antioxidant capacity. The spectral analysis results showed that the antioxidant capacity was positively related to quinic acid, bergapten, oxypeucedanin, phellopterin, isoimperatorin and falcarinolone. CONCLUSION In this study, a quality evaluation model of Angelica dahurica cv. Qibaizhi based on chemical composition and activity is established, which provides a reference for elucidating the antioxidant active ingredients and quality control in Angelica dahurica cv.Qibaizhi.

Hydrogels and nanoparticles have great application prospects in drug delivery. Hydrogels possess good biocompatibility and physicochemical versatility to enable disease-triggered in situ self-assemble and sustained or stimuli-responsive drug release. The high targeting ability and low toxicity of nanoparticles can significantly improve the delivery efficiency of antitumor drugs. However, hydrogels and nanoparticles still face many challenges in their applications, for example, hydrogels suffer from low mechanical strength and have difficulty in delivering hydrophobic drugs, while nanoparticles have off-target effects and low accumulation and retention in tumors. Therefore, the incorporation of nanoparticles into the hydrogel network can form a novel multifunctional system that enables the hydrogel to serve as a reservoir for the local delivery of nano-drugs into tumors, thereby combining the advantages of two preparations to produce synergistic therapeutic effects. Herein, we review the design of drug release behavior of nanoparticle-hydrogel composite systems, and outline the recent progress of nanoparticle-hydrogel composite systems in the local application of antitumor drugs, including intratumoral and peritumoral injections, subcutaneous or intramuscular injections, transdermal administration and intraluminal administration, providing insights and references for the rational design of novel anti-tumor dosage forms and preparations.

OBJECTIVE To establish the fingerprints of 50 batches of Arnebiae Radix and the methods for determination of the contents of six components. METHODS A high performance liquid chromatographic (HPLC) method was used for the determination of 50 batches of Arnebiae Radix on an Agilent 5 TC-C₁₈(2) column, with acetonitrile-0.05% formic acid aqueous solution as the mobile phase, gradient elution at a flow rate of 1 mL·min^-1, detection wavelength of 275 nm, and column temperature of 30 ℃. The contents of six naphthoquinone components, i.e., alkannin, acetylshikonin, deoxyshikonin, β-acetoxyisovalerylshikonin, isobutyryl alkannin, and β,β'-dimethylacrylalkannin were determined. The fingerprints of the herbs were established by HPLC fingerprinting combined with the “Chinese Medicine Chromatographic Fingerprint Similarity Evaluation System (Version 2012A)” to evaluate the similarity and confirm the common peaks. RESULTS The methodological validation results showed that the peak areas of the six components had good linear relationships in the corresponding concentration ranges, and the contents of the six components ranged from 0.006%-0.237%, 0.009%-1.759%, 0.001%-0.129%, 0.002%-0.625%, 0.011%-0.609%, and 0.016%-1.712%, respectively. The HPLC fingerprints of Arnebiae Radix were established, and a total of nine common peaks were calibrated. The similarity fitting was performed by using the TCM fingerprinting software, and it was found that the similarity of the 50 batches of Arnebiae Radix varied greatly. CONCLUSION The HPLC fingerprint of Arnebiae Radix established in this study and the method of simultaneous determination of the contents of six components are simple, stable, accurate, reliable, and reproducible, which can provide a basis for the quality study of Arnebiae Radix.

OBJECTIVE To study the identification of Sargassum fusiforme and other sargassos (algae from the genus Sargassum) by targeted lipidomics combined with chemotaxonomy methods. METHODS Twenty-seven batches of S. fusiforme were collected and the GC-MS fingerprint of fatty acids (FAs) was established. The principal component analysis and orthogonal partial least squares discriminant analysis were used to compare S. fusiforme with other 8 species, total 22 batches of sargassos. Based on the differential FAs between S. fusiforme and other sargassos, combined with the analysis of FA biosynthesis pathway, the Q-marker was screened. RESULTS A total of 29 common FAs were identified by the fingerprint, and the similarity was greater than 0.96. The chemical pattern recognition method can well distinguish S. fusiforme from other 8 species, and the discrimination model established by this method can accurately verify 15 batches of commercial slices. Based on the analysis of differential FAs and FA biosynthesis pathway, monounsaturated fatty acid was found to play an important role in chemical classification. The ratio C20∶1 n-9/C16∶1 n-7 was proposed to be used as a candidate Q-marker for the identification of S. fusiforme and 14 batches of commercial slices can be accurately verified. CONCLUSION This study establishes the identification model of S. fusiforme and other confused sargassos, finds the Q-marker of FAs, and provides a reference for the establishment of the identification method of TCM SARGASSUM.

OBJECTIVE To investigate the protective effect and mechanism of salvianolic acid A (SAL-A) on mouse brain microvascular endothelial (b.End.3) cell apoptosis and oxidative stress injury after oxygen-glucose deprivation/reoxygenation (OGD/R). METHODS OGD/R injury was established with b. End.3 cells for injury model. Cell apoptosis was detected by Annexin V-fluorescein isothiocyanate (Annexin V-FITC) probe and propidium iodide (PI) staining; Reactive oxygen species (ROS) generation was detected by 2,7-dichlorofluorescein diacetate (DCFH-DA) probe; Mitochondrial membrane potential was detected by JC-1 staining. The expression of proteins related to apoptosis, oxidative stress, and Akt/GSK3β/Nrf2 pathway was detected by Western Blot. RESULTS Compared with the control group, the expression of anti-apoptotic proteins, mitochondrial membrane potential, expression of related antioxidant proteins and Akt/GSK3β/Nrf2 pathway were significantly decreased in the OGD/R group (P<0.05). In addition, the number of apoptotic cells, the expression of apoptosis-inducing proteins, and the production of intracellular ROS were significantly increased in the OGD/R group (P<0.05). Whereas, the SAL-A treatment can improve OGD/R-induced cell apoptosis and oxidative stress, the expression of Akt/GSK3β/Nrf2 pathway were significantly increased in the SAL-A-10 group. CONCLUSION SAL-A inhibited OGD/R-induced apoptosis and oxidative stress injury in b.End.3 cells, and the mechanism may be related with affecting the Akt/GSK3β/Nrf2 pathway.