The hippocampal proteomics mechanism of thalidomide in Alzheimer's disease model mice induced by Aβ / 中国药理学通报

Aim To observe the effect of thalidomide on the learning and memory ability and hippocampal tissue proteome of Alzheimer's disease(AD)mice,to screen the differential proteins of thalidomide in preventing and treating AD,the pathways involved in regulation,and to explore its possible mechanism. Methods The experimental mice were randomly divided into blank control group,model group,and thalidomide high and low dose groups. The drugs were administered by gavage every day for 21 days. After the administration,Morris water maze test was used to evaluate the learning and memory abilities of the mice,HE staining and Nissl staining were used to observe the pathological tissue morphology of the mouse hippocampus,ELISA was employed to detect the mitochondrial respiratory chain enzyme complex in mouse brain,and the Label-free proteomics method was used to screen different groups of hippocampal proteome proteins. Results The results of the Morris water maze showed that compared with the model group,the escape latency time of the drug group was significantly reduced,and the number of crossing the platform significantly increased(P<0.05). Thalidomide administration could improve the morphological structure of neurons in hippocampus,and could increase the activity of the mitochondrial respiratory chain complex ,Ⅱ, and of the brain tissues of AD mice(P<0.05). A total of 4 378 differential proteins were identified,which had a significant regulatory effect on the expression of 580 proteins in hippocampus of AD mice(P<0.05). Energy metabolism may jointly participate in the regulation of neurodegeneration pathways-changes in pathways such as various diseases and Alzheimer's disease. Conclusions Thalidomide can significantly improve the learning and memory function of AD model mice induced by Aβ

Long-term intermittent fasting induces abnormal lipid accumulation in mouse liver / 生理学报

Short-term intermittent fasting (IF) is beneficial to weight control in patients with nonalcoholic fatty liver disease, but the impact of long-term IF is not clear. In this study, healthy C57BL/6N mice with 4-month alternate day fasting (ADF) were used to study the effects of long-term IF on systemic and liver lipid metabolism. The results showed that, compared with the Ad Libitum group, the weight and food conversion rate of mice in the ADF group were markedly decreased and increased respectively, and the liver index and the liver content of triglyceride were significantly increased by pathological examination. qRT-PCR analysis revealed that the mRNA expression of the lipogenesis gene Pparγ and lipolysis gene Atgl was up-regulated in the ADF group (P < 0.05). Western blot analysis showed that the ratio of microtubule associated protein LC3-II/LC3-I was increased, while the abundance of autophagy adaptor protein p62 was decreased in the ADF group. In addition, autophagy signal positive regulation key factor AMPK phosphorylation was increased (P < 0.05), and negative regulation factor mTOR phosphorylation was decreased (P < 0.05) in the ADF group, indicating that hepatocyte autophagy activity was elevated. Taken together, ADF for 4 months results in an excessive liver triglyceride accumulation, accompanied by a marked decrease in liver mTOR phosphorylation and a significant increase in hepatic autophagy.

Research progress in antitumor mechanism of metformin independent of adenosine monophosphate activated protein kinase pathway / 中国药理学与毒理学杂志

Metformin is an oral hypoglycemic drug widely used in the treatment of type 2 diabetes mellitus. Epidemiological studies have showed that metformin is deeply involved in the prevention and treatment of cancer. And the antitumor mechanism of the effect of metformin has become a hot spot under intensive study. Most studies suggest that the pharmacological effect of metformin on cancer depends on the activation of adenosine monophosphate activated protein kinase (AMPK) pathway. Recently, some researchers proposed the anti-tumor mechanism of metformin through the AMPK independent pathway. Metformin affects the growth, invasion and migration of tumor cells by improving insulin sensitivity, down-regulating Ras-related GTP-binding protein Rag activity and the expressions of DNA damage response regulator gene 1 (Reddl), inhibiting the mammalian target of Rapamycin signaling pathways, and reducing the activity of Ras-related C3 botulinum toxin substrate and the protein expression of cell division cycle 42 and matrix metalloproteinases. In this paper, the antitumor effect and related mechanism of metformin independent of AMPK pathway are summarized, which offers a new perspective from which to explore the antitumor effect of metformin.

Research progress on Q-biomarker of Chinese materia medica / 中草药

Quality marker (Q-marker) is one of the new directions in the research of quality control of Chinese materia medica. The Q-marker can be either quality chemomarker (Q-chemomarker) or quality biomarker (Q-biomarker). The establishment of a good Q-biomarker can improve the linkage of Chinese medicine quality control methods and standards with clinical safety and effectiveness, at the same time considering the operability of quality control. At present, the FDA of China and the United States have adopted biological evaluation methods as an important part of quality control of Chinese medicines and botanicals medicines. Many companies and researchers are also actively exploring and developing methods for the evaluation of quality of Chinese materia medica. This paper further enlarges and expands the methods and indicators of biological evaluation of Chinese materia medica, and summarizes and analyzes relevant research examples from the perspective of Q-biomarker. We hope that it would promote the richness and development of the theoretical system of Chinese medicine quality markers, so as to provide a reference to solve the practical problems for the quality control and evaluation of Chinese materia medica.

Scientific research of laxative action of Cannabis Fructus based on intestinal environment adjustment / 中草药

Objective: To investigate the effects of the water extract of Cannabis Fructus on intestinal flora structure, short chain fatty acids (SCFAs) levels, and pathological changes of colonic inflammation, and explore its scientific connotation of laxative action. Methods: A total of 48 Wistar rats (SPF grade), sex in half, were randomly divided into control group, model group, Cannabis Fructus water extract group, and positive control group. The rats of model group were caused constipation and enteric flora disturbance by diphenoxylate. The bacterial counts in the fresh stool were tested by plate count method 28 days after administration, and the changes of intestinal flora structure in cecum was detected by PCR-DGGE method, the contents of SCFAs in cecum were determinated by GC method, and the coton pathological score of the rats were studied by HE staining method. Results: Compared with the blank group, the number of Escherichia coli and Bifidobacterium was increased but Bacillus acidi lactici was decreased significantly (P < 0.05, 0.01) in fresh stool of the model group. The ratio of Firmicutes bacteria to Bacteroides bacteria was increased, the number of Butyricicoccus and Cillobacterium was decreased, the concentration of acetic acid and butyric acid was increased significantly (P < 0.05, 0.01), and the pathology score of colon was increased significantly (P < 0.01). Compared with the model group, Cannabis Fructus water extract can adjust the ratio of Firmicutes bacteria to Bacteroides bacteria, increase the number of Cillobacterium, Butyricicoccus, Blautia, and Oscillibacter in cecum, decrease the number of Clostridium IV, etc., decrease the concentration of acetic acid and butyric acid (P < 0.05), reduce the pathology score, and improve the damage of intestinal structure (P < 0.05). Conclusion: Cannabis Fructus water extract can alleviate constipation and accelerate the recovery of colitis injury in model rats. The functional route of Cannabis Fructus water extract may relate to the disorder of intestinal flora balance adjustment and effect of the concentration of SCFAs so as to change the acidic environment in the intestinal tract.

Advance in research on cell-derived nanomedicine delivery system / 药学学报

Nanoparticles hold great potential in the improvement of the therapeutic activities of many drugs. Synthetic approaches are dominant in the conventional approach for nanoparticles design and engineering strategies. However, combination of synthetic nanoparticles with natural biomaterials have recently gained much attention. By taking inspiration from nature, cell-derived nanomedicine delivery system has been created, which is a biomimetic platform consisting of a nanoparticulate core coated with cell or cell membrane. Compared to the conventional drug delivery systems, this novel system combines the unique functionalities of cells and engineering versatility of synthetic nanomaterials for effective delivery of therapeutic agents. With existing of cell, nanomedicine has significantly improved the biocompatibility, accurate delivery and long half-life in circulation as well as reduced the toxicity and side effect of drugs. Moreover, the delivery system can interact with the incredibly complex biological systems that exist within the body, such as actively targeting the inflammatory sites and tumors. Hence, it can be applied to drug delivery, tumor radiotherapy, and vaccine preparation. The cell-derived nanomedicine delivery system emerging as a novel delivery strategy, have the potential to significantly advance the nanomedicine to improve the therapeutic efficacy. The recent research in characteristics, preparation and application of erythrocyte, mononuclear phagocyte, bacteria and tumor cell as nanomedicine delivery carrier are reviewed.

History and development of biological assay for quality of chinese materia medica / 中草药

Biological assay is an important method for quality control of drugs. There are limitations in the quality control system of Chinese materia medica (CMM) based on physico-chemical analysis methods due to the complicated chemical constituents and multifold pharmacodynamics of CMM. Recently, the application of biological assays to CMM attracted much attention and application. From the historical development perspective, the idea and method of biological assay for the quality control of CMM have existed since the ancient times. Especially in recent years, the introduction of biological assay to botanical drugs and CMM submitted in new drug applications were emphasized by both the U. S. and Chinese Food and Drug Administration. This means that a common view has been formed that using biological assay in the quality control of CMM and botanical drugs is proper. Now, a new era of biological assay for the quality control of CMM has come. Based on the review and expectation of biological assay for the quality control of CMM, we creatively put forward an integrated quality concept for CMM which takes biological assay as core.

Screening bioactive compounds inhibiting influenza virus from isatidis radix by ultrafiltration mass spectrometry / 中国中药杂志

In vitro neuraminidase inhibition assays and ultrafiltration liquid chromatography with diodearray detector coupled to time of flight mass spectrometer (UPLC-DAD-TOF-MS) were combined to screen bioactive compounds inhibiting neuraminidase from Isatidis Radix. By comparing the compounds from Isatidis Radix before and after ultrafiltration, we found that arginine, goitrin and adenosinea can bind with neuraminidase, and the binding degree of the three compounds were (36.23 +/- 1.12)%, (32.54 +/- 1.02)% and (9.38 +/- 0.47)%, respectively. The IC50 of arginine and goitrin were (1.16 +/- 0.02), (1.20 +/- 0.02) g x L(-1), respectively. While the IC50 of adenosinea was higher than 500 g x L(-1). The results showed that arginine and goitrin might be the main compounds with antiviral activity of Isatidis Radix. This study may provide a useful method for the screening of bioactive compounds and quality control of Isatidis Radix.

Bio-evaluation methods and optimization for Isatidis Radix quality control based on antiviral activity detection / 中草药

Objective: To establish and optimize a bio-evaluation method based on antiviral activity detection for Isatidis Radix quality control. Methods: According to the fact that antivirus activity is the main pharmacological effect of Isatidis Radix, the means of hemagglutination activity detection and the means of influenza virus neuraminidase (NA) activity detection were used for establishing two antiviral activity detection methods for the bio-evaluation of Isatidis Radix. In addition, contrastive analyzation and optimization of the two methods were carried out in this study. Results: The two methods could be used to evaluate and distinct the biological activities of different batches of Isatidis Radix samples and both methods have good repeatabilities (RSD = 7.0% and 5.78%). The result shows a good correlation (P<0.01, r = -0.81) between the neuraminidase inhibitory activity and the hemagglutination activity of Isatidis Radix. The relationship between agglutination activity and antiviral effect of Isatidis Radix was further verified. Agglutination detection method has the superiority to its safety, inexpensiveness, easiness, and practicality. Conclusion: This novel method could reflect the pharmacodynamic activity of Isatidis Radix on anti-influenza virus and be helpful for the quality control of Isatidis Radix. This study provides reference for the bio-evaluation on the quality control of Chinese materia medica.

Hemagglutination activity of radix isatidis detected by microcalorimetry / 药学学报

In this study, microcalorimetry was adopted to establish a novel method for detecting the hemagglutination process of Radix Isatidis (Banlangen in Chinese, BLG), and to evaluate the hemagglutination activity diversity of BLG from various habitats. The hemagglutination biothermokinetics curves of positive reagent (phytohemagglutinin, PHA) and 8 batches BLG from different regions of the hemagglutination with 20% rabbit erythrocyte were recorded by microcalorimetry, then biothermokinetics parameters were abstracted, the hemagglutination utility of samples were calculated and analyzed by principal component analysis (PCA) and cluster analysis (CA), meanwhile the results were authenticated by micro-plate agglutination. It showed that the hemagglutination was an exothermic reaction, the reaction rate constant (k: 0.039-73.6 min(-1)), maximum reaction power (Pmax: -1 140.2 - 988.2 microW) and reaction enthalpy (Hi: -529.9 - 717.9 microJ) had good linear correlation with BLG extraction concentration (0.2-1.0 g mL(-1), r > 0.97), and PCA showed Pmax (531-1 335 microW) and Ht (585.2-989.2 microJ) could represent the hemagglutination activity diversity of BLG samples, just confirming with the results of micro-plate agglutination (the agglutination dilution was 3-11 respectively). According to the hemagglutination utility, the BLG samples from Good Agriculture Practice (GAP) regions, main producing area and general regions could be clustered correctly; meanwhile, the biothermokinetics curves with perfect distinctive fingerprint and specificity could give out more information for the quality control and evaluation for BLG. In conclusion, the microcalorimetry method established for detecting the hemagglutination activity of BLG samples on rabbit erythrocyte is sensitive and reliable, and could be adopted as an effective technique in detection aggulatination precisely, quantitatively and consecutively; and provide a novel approach for examining and evaluating quality for Chinese herbal medicine with aggulatinative activity such as BLG.

Agglutinated activity bioassay method for the determination of antivirus potency of Banlangen granula / 药学学报

To establish a bioassay method and quality standard of Banlangen granula, agglutinated activity assay was used in the analysis of the traditional Chinese medicine, Banlangen granula. It showed that masculined effect could be picked up effectively and the products quality of different pharmaceutical factories and different batch numbers from the same factory could be revealed conveniently, accurately, quickly and directly with this method (valence value was between 2 and 11). The established bioassay method had a good reproducibility with RSD = 2%. The dependablity of the activity of red cell agglutination and restrainting influenza virus NA was conspicuous (r2 = 0.878 3). In conclusion, this bioassay method is suitable to control and evaluate the quality of Banlangen granula. Thus the method may provide a simple and effective technique in supervising and examining the quality of other traditional Chinese medicine.

Determination of colistin based on biothermal activity detection method / 药学学报

A biothermal activity detection method has been established to determine the potency of colistin. The biothermal activity fingerprints of E. coli with colistin were determined. There was a good linear relationship (r = 0.993) between logarithm concentration of colistin (lgC) and lag rate of growing time (Deltat%) when the concentrations of colistin ranged from 17.0 to 41.6 u x mL(-1). The average recovery rate was 100.3% (n = 9). Using this method, there was no significant difference between results of colistin potency measurement and those using cup-plate method (P > 0.05). As a result, biothermal activity detection method is sensitive, accurate, rapid, convenient and feasible to determine the potency of colistin. This method can also be applied in real time and online to monitor the process of bacterial growth and could be complementary to the cup-plate method.

Biological evaluation of Radix Isatidis based on neuraminidase activity assay / 药学学报

Radix Isatidis (Banlangen in Chinese) is a traditional Chinese medicinal (TCM) herb, and is frequently used for treating influenza. However, the current quality control method for Radix Isatidis should be developed since it has little correlation to the pharmacodynamic action. In this paper, the in vitro inhibitory action of Radix Isatidis on neuraminidase (NA) was investigated by fluorometric assay with 4-methylumbelliferyl-D-N-acetylneuraminate (FL-MU-NANA) method. Based on the method, the experimental condition was optimized and a bioassay statistic method was established according to the reaction type and the regularity of "parallel lines of qualitative effect". Then the bioassay method of Radix Isatidis was established. This study indicated that Radix Isatidis had obvious in vitro inhibitory activity on NA with IC50 = (0.90 +/- 0.20) mg x mL(-1) (herb). The correlation between logarithmic dose and reaction rate showed an "S" shape--is quite similar to Tamiflu's reaction curve, which hinted that Radix Isatidis had the same inhibitory function on NA as Tamiflu. The established bioassay method of "parallel lines of qualitative effect" had a good reproducibility (RSD = 5.78%). The results of potency determination of Radix Isatidis were reliable (reliability test: deviation from straight line P > 0.05, deviation from parallel line P > 0.05) and well regular. As a conclusion, this bioassay method is suitable to control and evaluate the quality of Radix Isatidis.

Essential issues of bioassay methods for quality control of laxative traditional Chinese medicines / 药学学报

The essential issues of bioassay methods for quality control of traditional Chinese medicines (TCM) were investigated and discussed through the instantiation of developing the bioassay methods for laxative drugs. For the relatively broad variation of the quality of TCM, which might be influenced much by many factors, the parallel lines model of quantitative response is preferred to control the quality of TCM for its relatively high accuracy. The parallel model of quantal response is alternative while the sample can not meet the reliability standard for quantitative response model. According to the requirement of homogeneity between reference and sample for bioassay, the extract from referenced crude meterial is suitable to be used as reference substance after standardizing and defining by chemical substances, and to give reference to the establishment and reproducibility of authorized standard substance. The results of determination of the purgative biopotency of different species of rhubarb and the compound preparations showed that the bioassay methods and self-made reference substance established in this study could be used to control the quality of laxative medicines.