An analytical method of ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC/PDA-QDa) for the qualitative and quantitative analysis of Δ9-tetrahydrocannabinol (Δ9-THC), cannabidiol (CBD), and cannabinol (CBN) in confiscated cannabis was developed.The seized cannabis was extracted in methanol by sonication.The binary mobile phase consisted of methanol (containing 0.1% formic acid) and water.After centrifugation, the supernatant was separated on Waters UPLC BEH C18 column (50 mm×2.1 mm, 1.7 μm) with isocratic elution at a flow rate of 0.2 mL/min.The three cannabinoids were analyzed by photodiode array (PDA) detector at 220 nm and confirmed by mass spectrometer QDa.The correlation coefficient of standard curve for the three cannabinoids in linearity range was not less than 0.999, as well as the recoveries were 82%-102% with the relative standard deviations (RSD) of 0.36%-4.12% at three spiked levels.The method is specific, easy, quick and suitable for confirmation of the cannabinoids in seized cannabis.Cannabis plants in different areas were classified by their chemical phenotype as drug-type or fiber-type plants, taking into account the phenotypic index Δ9-THC, (Δ9-THC+CBN)/CBD, or the Δ9-THC/CBD and the (Δ9-THC+CBN)/CBD ratios.The analysis of the original composition of plant material is necessary for the detection and the quality control of cannabis plants.

Objective To express a recombinant protein TFPR1 ( the functional region of the snake venom proteins from Trimeresurus flavoviridis) in Pichia pastoris expression system. Methods The target gene was codon-optimized and synthesized according to the sequence of the conserved structural do-main of triflin and then cloned into the Pichia pastoris expression vector pPICZαA to construct the recombi-nant expression plasmid pPICZαA-TFPR1. The recombinant plasmid pPICZαA-TFPR1 was electroporated into the yeast strain X33. The transformed strains carrying expression plasmid were screened out with Zeocin and then induced by methanol to express the recombinant protein TFPR1. ELISA was performed for the screening of positive clones. SDS-PAGE and Western blot were used for further identification of the ex-pressed products. Results The recombinant plasmid pPICZαA-TFPR1 was successfully constructed. The recombinant protein TFPR1 was expressed in a secreted form at a molecular weight of 16×103. Conclusion The recombinant protein TFPR1 was successfully expressed in Pichia pastoris expression system, which laid a foundation for further researches on its biological function and application as an adjuvant.

Objective To explore the medication regularity of prescriptions in Universal Relief:Cough Medicine in that to provide worthy reference for clinical treatment of cough.Methods The prescriptions in Universal Relief:Cough Medicine was collected and then established the database using Excel 2016 softwore,and carried out medication frequency analysis.SPSS Modeler 18.0 and Lantern 5.0 were used to perform association rules analysis and latent structure model analysis to explore medication rules of prescriptions in Universal Relief:Cough Medicine.Results 607 prescriptions were collected,involving 335 traditional Chinese medicines.A total of medication frequency is 3941,The most frequently used TCMs were ganca(glycyrrhizae radixet rhizoma)and shengjiang(zinglberis rhizoma recens)and kuxingren(armeniaeae semen amarum).The main efficacy of the medicine is to dissipate phlegm,relieve cough,relieve asthma,and replenish deficiency,and warm in nature,spicy,bitter,and sweet in taste and lung and spleen in meridian.14 strongly correlated material rules were sorted out by association rule analysis such as banxia(pinelliae rhizoma)+gancao(glycyrrhizae radixet rhizoma)→shengjiang(zinglberis rhizoma recens)、banxia(pinelliae rhizoma)→shengjiang(zinglberis rhizoma recens)and mahuang(ephedrae herba)+gancao(glycyrrhizae radixet rhizoma)→kuxingren(armeniaeae semen amarum).12 variables were obtained by latent structure model analysis,and according to professional knowledge inference,3 integrated cluster models and 1 single cluster and 4 core prescriptions were obtained.It was inferred that the common syndromes of Universal Relief:Cough Medicine were cold drink stoping in the lung,qi deficiency external phlegm dampness inside,phlegm heat blocking lung,and lung deficiency prolonged cough.Conclusion The medication rules of prescriptions in Universal Relief:Cough Medicine is mainly benefiting qi and resolving phlegm,cure both the symptoms and the disease,and in different situations combine with different treatment methods,such as warming the lung and resolving phlegm,clearing heat and resolving phlegm,tonifying qi and resolving external symptoms,and astringent lung qi.

【Objective】 To compare the histological characteristics of porcine pancreatic elastase （PPE） induced abdominal aortic aneurysms （AAA） and angiotensin Ⅱ （AngⅡ） induced AAA in mice. 【Methods】 In the PPE group， the mouse abdominal aorta segment from the infrarenal abdominal aorta to the iliac artery was isolated and its branch arteries were ligated to avoid leakage during PPE perfusion. We perfused the isolated aorta segment with a PPE solution at a concentration of 1.5 U/mL for 5 min and then closed the abdominal cavity. The diameter of the abdominal aorta was measured before and 14 days after the surgery， and the perfusion segment of the arteries was collected at day 14 after the surgery. The histological characteristics of the aneurysm were analyzed and graded by histological and immunohistochemical methods. In the AngⅡ group， ten apolipoprotein E knockout mice were prepared， and AngⅡ [1 000 ng/(kg·min)] was infused with osmotic pumps for 28 days. The aorta was separated and the aneurysm aorta segment was analyzed. The wild type mice were used as normal health controls. 【Results】 In the PPE group， the diameter of the PPE perfused aorta segments increased and was significantly larger than the basal diameter ［（0.52±0.02） mm vs. （1.23±0.11） mm］ at day 14 after surgery. All the ten mice developed AAA after PPE application. The histological results showed typical pathological features of AAA in PPE perfused mice， such as elastic fiber breakage， smooth muscle exhaustion， and increased inflammation. Six of the ten mice developed aneurysms after AngⅡ infusion （6/10）. The aneurysms/dilatations were mostly in the suprarenal abdominal aorta， but also in the thoracic aorta and aortic arch. The histology analysis showed that the formation of arterial dissection was common after AngⅡ infusion， and the typical vascular “false lumen” was found. The breakage of elastic fibers， the exhaustion of smooth muscle damage， and the inflammatory response were not as typical as the PPE model in AngⅡ perfused animals. 【Conclusion】 The histological characteristics of PPE induced AAA are very typical and well present the inflammatory process in the development of aneurysm. The AngⅡ model is suitable for the study of aneurysms combined with aortic dissection. Both models have their own advantages and can complement each other.