OBJECTIVE: To establish a preparation process of pivotal intermediate of SKI2496, which is low-cost, environmental-friendly and suitable for industrialization as well. METHODS: 1-(2-Fluoro-6-(trifloromethyl)benzyl)urea(2) was synthesized from 2-fluoro-6-(trifluoromethyl)benzylamine(1) with urea,followed by aminolysis with t-butyl acetoacetate and cyclization to give 1--6-methylpyrimidine-2,4(1H,3H)-dione(4).Finally,the title product was obtained via bromation and condensation reaction with piperazine. RESULTS: The synthetic process included four steps with an overall yield of 44.6%(based on compound 1) and its structure was confirmed by 1H-NMR and MS. CONCLUSION: The process is easy to operate, safe and suitable for industrial production.

Qigui Tongfeng tablet (QLTFT) is a traditional Chinese medicine with good effect for treating gout. Here, network pharmacology method and molecular similarity analysis were utilized to study the effective substance basis and molecular mechanism of the QLTFT on the gout. The similarity to the medicinal compounds is reflected in the Tanimoto coefficient that gives the structural similarity of two compounds. Operationally, similar modifiers were described as pairs of concepts with a similarity score of 0. 500. The results of the molecular similarity analysis suggested that the flavonoids in QLTFT could be new leads for gout. Furthermore, complex biological systems may be represented and analyzed as computable networks. Two important properties of a network were degree and betweenness. Nodes with high degree or high betweenness may play important roles in the overall composition of a network. And the results of network analysis showed that dongbeinine, verticinone-N-oxide, verticine N-oxide, peimine, peiminine, isobaimonidine, dongbeirine, peimisine and simi-arenol which with high degree acted on xanthine dehydrogenase/oxidase, matrix metalloproteinase-9, an arachidonate 5-lipoxygenase-activating protein, tyrosine-protein kinase and etc. Inhibition of these targets can prevent the formation of uric acid, reduce inflammation by uric acid and regulate the body's immune response. Thus, these compounds may be the main effective substance basis. The research results not only reveals its molecular mechanism, but also provide a theoretical basis for the quality control of drugs and clinical application.
Gout ; drug therapy ; Humans ; Medicine, Chinese Traditional ; Pharmacology ; methods ; Tablets ; Technology, Pharmaceutical ; methods

Objective: To observe the changes in numbers and degranulation status of local mast cells (MCs) of the complementary acupionts, as well as the concentrations of serum histamine and bradykinin after acupuncture at rat's complementary acupionts of Yanglingquan (GB 34) and Yinlingquan (SP 10), for exploring the mechanisms of the synergistic effect of complementary acupionts. Methods: Using random number table method, 40 Wistar rats were randomly divided into a blank control group (group K), an acupuncture at Yanglingquan (GB 34) group (group A), an acupuncture at Yinlingquan (SP 9) group (group B), an acupuncture at Yanglingquan (GB 34) and Yinlingquan (SP 9) group (group AB), and an acupuncture at points near Yanglingquan (GB 34) and Yinlingquan (SP 9) group (group CD), 8 rats in each group. Group A received acupuncture at Yanglingquan (GB 34); group B received acupuncture at Yinlingquan (SP 9); group AB received acupuncture at Yanglingquan (GB 34) and Yinlingquan (SP 9); group CD received acupuncture at the control points [points 3 mm away from Yanglingquan (GB 34) and Yinlingquan (SP 9) respectively]. Acupuncture was performed at bilateral points in each group. Rats in group K were fixed using the same method as rats in the other 4 groups without acupuncture stimulation. Needles of 0.35 mm in diameter and 40 mm in length were used for acupuncture. Needle handle was connected to the G6805-Ⅱ mode electroacupuncture (EA) device after needling qi was obtained, with sparse-dense wave, frequency of 2 Hz/100 Hz and current of 1 mA to keep the needle handles slightly tremulous while the rats kept quiet. Rats were continuously stimulated by EA for 20 min each time. Experimental interventions were conducted on the 1st, 3rd, 5th and 7th days when the experiment started, for 4 times in total. Specimens from rats in each group were collected 2 h after the 7th day. The levels of histamine and bradykinin in serum were detected by enzyme-linked immunosorbent assay (ELISA). Local tissues of the points were used to prepare cryosections. The changes of MCs were observed after toluidine blue staining. Results: Compared with group K, the numbers and degranulation rates of MCs in group A, group B, group AB and group CD were significantly increased (all P<0.05). The number and degranulation rate of MCs in group AB were significantly higher than those in group A, group B and group CD (all P<0.05). The order of histamine levels from high to low was: group AB >group B > group A > group CD > group K, and the differences were statistically significant (all P<0.05); the levels of bradykinin in group AB, group A and group B were significantly higher than those in group K and group CD (all P<0.05). There was no significant difference in the bradykinin level between group A and group B, nor between group CD and group K (all P>0.05). Conclusion: The number and degranulation rate of MCs of the complementary acupionts are significantly increased after acupuncture at complementary acupionts of Yanglingquan (GB 34) and Yinlingquan (SP 9) of rats, and the serum levels of histamine and bradykinin are increased, which may be one of the mechanisms of the synergism produced by the combination of complementary acupionts.

Education, Dental ; Oral Medicine ; education

To study the in vitro anti-angiogenesis effect of three curcumin pigments (curcumin, demethoxycurcumin, bisdemethoxycurcumin). In the study, the inhibitory effect of the three curcumin pigments on proliferation of HUVEC cells induced by OX-LDL and the effect on migration of HUVEC cells were detected. The effect on neovascularization was observed by chorioallantoic membrane (CAM) test. The effect on cell adhesion factors ICAM-1 and VCAM-1 of HUVECs were tested by Real-time RT-PCR. It was found that the three curcumins could inhibit the proliferation of HUVEC cells induced by OX-LDL within the dosage range 4, 8, 16 mg x L(-1), with a dose-dependence. The proliferative effect of curcumins on HUVECs was greater than the other two derivatives (P < 0.01). All of the three curcumin pigments inhibited the migration of HUVEC cells and the angiogenesis of chick chorioallantoic membrane (CAM). The migration inhibition rate of curcumins at middle and high concentrations was greater than the other two (P < 0.01). All of the three curcumin could down-regulate the expression of VEGF and ICAM-1, and curcumins showed more obvious effect in down-regulating VEGF than demethoxycurcumin and bisdemethoxycurcumin(P < 0.01); Bisdemethoxycurcumin showed the most significant effect in down-regulating ICAM-1 (P < 0.01). All of the three showed no remarkable effect on expression of VCAM-1, and only bisdemethoxycurcumin showed the down-regulating effect (P < 0.05). According to the findings, all of the three curcumin pigments could resist angiogenesis by inhibiting proliferation and migration of endothelial cells and down-regulating the expression of VEGF and adhesion molecules ICAM-1.
Angiogenesis Inhibitors ; pharmacology ; Animals ; Cell Movement ; drug effects ; Cells, Cultured ; Chorioallantoic Membrane ; drug effects ; Curcumin ; analogs & derivatives ; pharmacology ; Humans ; Intercellular Adhesion Molecule-1 ; genetics ; RNA, Messenger ; analysis ; Vascular Cell Adhesion Molecule-1 ; genetics ; Vascular Endothelial Growth Factor A ; genetics

ObjectiveTo observe the protective effect of extract from Morindae officinalis on the oxidative injury of human sperm motility.MethodsThe sperm specimens were cultivated in vitro, the reactive oxygen species (ROS) was made from hypoxanthine-xanzine xanzine (HX-XO), and sperm specimens were co-incubated with ROS and different dosages of extract and oxygen. The motor parameters were evaluated, the motility were analyzed by the test of capillary tube penetrating, and compared with the control group.ResultsThe sperm function were injured, the motility and penetrating ability decreased evidently under the action of ROS, and with a significant difference compared with the normal group ( P<0.001). The sperm motility and penetrating ability could be improved by all of the small, medium and massive extract compared with vitamin C, especially 0.25 and 0.5 g/ml Morindae officinalis had more evident action in the rate of progressive motility and vitality exponent.ConclusionThe ROS can injury the sperm motility and penetrating ability, the appropriate content of extract from Morindae officinalis can significantly intervene peroxidation in sperm by the ROS, and protect the sperm motility.

(3S)-Linalool synthase (LIS) is a key enzyme involved in the monoterpene biosynthetic pathway. Based on our previous transcriptome study, the expression level of LIS gene was exceedingly related to glycyrrhizic acid (GA) biosynthesis. Therefore, we used hairy root culturing to further investigate the effect of LIS on the GA biosynthesis. A LIS gene (GenBank accession number: MZ169552) was cloned from Glycyrrhiza uralensis. The plant binary overexpression vector pCA-LIS was constructed by gene fusion. G. uralensis hairy roots overexpressing LIS were induced by the Agrobacterium rhizogenes ATCC15834. The expression levels of LIS were analyzed by real-time quantitative PCR (RT-qPCR) and the contents of GA in hairy root lines were determined by UPLC. It was found that in the hairy root lines overexpressing LIS, the expression levels of LIS were significantly higher than that in the wild type, while the contents of GA were remarkably lower than those in the wild type and negative control. These findings indicate that the expression level of LIS is negatively correlated with the accumulation of GA. In this study, LIS was cloned from G. uralensis for the first time and the negative regulatory effect of LIS on GA biosynthesis was confirmed by reverse genetics. This work provides support for further improvement of the molecular regulatory network of GA biosynthesis in G. uralensis.

AIMTo study chemical constituents from pine cone of Pinus armandii Franch.

METHODSThe constituents were isolated by chromatographic method and the structures were identified on the basis of spectral analysis.

RESULTSFour compounds were identified as 7-oxo-12alpha, 13beta-dihydroxyabiet-8(14)-en18-oic acid (I), 7-oxo-13beta-hydroxyabiet-8 (14)-en-18-oic acid (II), 8 (14)-podocarpen-13-on-18-oic acid (III) and lambertianic acid (IV).

CONCLUSIONCompound I is a new diterpenoid and compounds II, III were isolated from this plant for the first time.

Carboxylic Acids ; chemistry ; isolation & purification ; Diterpenes ; chemistry ; isolation & purification ; Fruit ; chemistry ; Molecular Conformation ; Naphthalenes ; chemistry ; isolation & purification ; Pinus ; chemistry ; Plants, Medicinal ; chemistry

Acetylcholine ; Adult ; Aged ; Coronary Vasospasm ; diagnosis ; Electrocardiography ; Humans ; Middle Aged ; Sensitivity and Specificity

OBJECTIVETo investigate association of mutation in WNK4 gene with essential hypertension and to analyze the expression of WNK4 gene.

METHODScSNP in WNK4 gene in a small samples was detected by sequencing, then PCR-RFLP was performed in 98 patients with essential hypertension and 95 control subjects. The expression profile of WNK4 gene was tested by RT-PCR.

RESULTSA cSNP was detected in WNK4 gene exon7 G1662A, and there were significant differences in the distribution of allele frequency of G1662A between essential hypertension group and control group. WNK4 gene were expressed in the tissues of kidney, brain, lung, heart, spleen and intestine of fetus.

CONCLUSIONWNK4 gene is well correlated with essential hypertension.

Adult ; Brain ; metabolism ; Female ; Gene Expression ; Gene Frequency ; Genes ; Genetic Predisposition to Disease ; Humans ; Hypertension ; genetics ; Kidney ; metabolism ; Lysine ; genetics ; Male ; Middle Aged ; Mutation ; Phenotype ; Protein-Serine-Threonine Kinases ; biosynthesis ; genetics ; RNA, Messenger ; biosynthesis ; genetics