Dietary supplementation with different alternative to in-feed antibiotic improves growth performance of broilers during specific phases.

The effects of substituting Bacillus subtilis, Astragalus membranaceus, and enzymes for aureomycin to improve the growth performance of broilers during specific phases were studied to develop alternatives to in-feed antibiotics and decrease drug residues in meat food and antibiotic resistance. Six hundred one-day-old broilers were randomly assigned to 5 groups. Broilers in the control group were supplied with basal diets (CT), and those in the remaining 4 groups were supplied with feed containing aureomycin premix (AU), B. subtilis powder (BS), A. membranaceus root powder (AM), and enzyme compound powder (EN), respectively. Compared to the control group, broilers in the other groups exhibited better growth performance during different phases. Microbial analysis of cecal contents suggested that treatment with BS or EN significantly increased the abundance of Lactobacillus or Bifidobacteria but inhibited Escherichia coli or Clostridium welchii; however, these bacteria were suppressed by AU treatment except C. welchii. The digestibility of the feed in vitro was significantly enhanced by adding BS or EN to the feed, consistent with findings for growth performance. In conclusion, dietary supplementation with 3 additives could improve the growth performance of broilers during specific phases. Future studies should focus on designing suitable schedules to partially replace in-feed antibiotics.

Detrusor underactivity influences the efficacy of TURP in patients with BPO.

PURPOSE: To investigate the effect of detrusor underactivity on the efficacy of TURP in patients with benign prostate obstruction. METHODS: A retrospective study of 350 patients with benign prostate obstruction who underwent TURP was carried out. Different degrees of bladder outlet obstruction were grouped by the bladder outlet obstruction index. ROC curves were used to calculate the optimal cut-off point for the bladder contractility index used to divide the DU patients into mild DU and severe DU patients. The effect of DU on the efficacy of TURP in benign prostate obstruction patients was studied by comparing the subjective and objective parameters preoperatively and 3 months postoperatively between severe DU, mild DU and non-DU benign prostate obstruction patients in two obstruction groups (20 ≤ BOOI < 40 and BOOI ≥ 40). RESULTS: According to the ROC curve, the optimal cut-off point for the bladder contractility index was 82; thus, 69 patients were considered mild DU patients (82 ≤ BCI < 100), 67 patients were considered severe DU patients (BCI < 82), and 214 patients were considered non-DU patients (BCI ≥ 100). Both the postoperative subjective and objective parameters of the non-DU, mild DU and severe DU patients significantly improved in two obstruction groups. However, in the 20 ≤ BOOI < 40 group, the successful improvement rates for the IPSS, IPSS-S, IPSS-V, QoL and fQmax in the severe DU patients were only 38.2%, 38.2%, 44.1%, 41.2% and 38.2%, respectively. CONCLUSION: Patients with varying degrees of benign prostate obstruction can benefit from TURP, but for patients with severe DU in the 20 ≤ BOOI < 40 group, TURP should be considered only after deliberation.

Crocetin: A Systematic Review.

Crocetin is an aglycone of crocin naturally occurring in saffron and produced in biological systems by hydrolysis of crocin as a bioactive metabolite. It is known to exist in several medicinal plants, the desiccative ripe fruit of the cape jasmine belonging to the Rubiaceae family, and stigmas of the saffron plant of the Iridaceae family. According to modern pharmacological investigations, crocetin possesses cardioprotective, hepatoprotective, neuroprotective, antidepressant, antiviral, anticancer, atherosclerotic, antidiabetic, and memory-enhancing properties. Although poor bioavailability hinders therapeutic applications, derivatization and formulation preparation technologies have broadened the application prospects for crocetin. To promote the research and development of crocetin, we summarized the distribution, preparation and production, total synthesis and derivatization technology, pharmacological activity, pharmacokinetics, drug safety, drug formulations, and preparation of crocetin.

Gardenia jasminoides Ellis: Ethnopharmacology, phytochemistry, and pharmacological and industrial applications of an important traditional Chinese medicine.

ETHNOPHARMACOLOGICAL RELEVANCE: Gardenia jasminoides Ellis is a popular shrub in the Rubiaceae family. The desiccative ripe fruits of this plant (called Zhizi in China) are well known and frequently used not only as an excellent natural colourant, but also as an important traditional medicine for the treatment of different diseases, such as reducing fire except vexed, clearing away heat evil, and cooling blood and eliminating stasis to activate blood circulation. It has also been declared as the first batch of dual-purpose plants used for food and medical functions in China. AIM OF THE STUDY: This review aims to provide a critical and systematic summary of the traditional uses, ethnopharmacology, phytochemistry, pharmacology, toxicity and industrial applications of Gardenia jasminoides Ellis and briefly proposes several suggestions for future application prospects. MATERIALS AND METHODS: The related information on Gardenia jasminoides Ellis was obtained from internationally recognized scientific databases through the Internet (PubMed, CNKI, Google Scholar, Baidu Scholar, Web of Science, Medline Plus, ACS, Elsevier and Flora of China) and libraries. RESULTS: Approximately 162 chemical compounds have been isolated and identified from this herb. Among them, iridoid glycosides and yellow pigment are generally considered the main bioactive and characteristic ingredients. Various pharmacological properties, such as a beneficial effect on the nervous, cardiovascular and digestive systems, hepatoprotective activity, antidepressant activity, and anti-inflammatory activity, were also validated in vitro and in vivo. Moreover, geniposide and genipin are the most important iridoid compounds isolated from Gardenia jasminoides Ellis, and genipin is the aglycone of geniposide. As the predominant active ingredient with a distinct pharmacological activity, genipin is also an outstanding biological crosslinking agent. Gardenia yellow pigment has also been widely used as an excellent natural dye-stuff. Hence, Gardenia jasminoides Ellis has been applied to many other fields, including the food industry, textile industry and chemical industry, in addition to its predominant medicinal uses. CONCLUSIONS: According to this review, Gardenia jasminoides Ellis is outstanding traditional medical plant used in medicine and food. Pharmacological investigations support the traditional use of this herb and may validate the folk medicinal use of Gardenia jasminoides Ellis to treat different diseases. Iridoid glycosides are potential medicines. Gardenia yellow pigment has been the most important source of a natural colourant for food, cloth and paint for thousands of years. This herb has made great contributions to human survival and development. Moreover, it has also achieved outstanding progress in human life and even in art. Although Gardenia jasminoides Ellis has extremely high and comprehensive utilization values, it is still far from being completely explored. Therefore, the comprehensive development of Gardenia jasminoides Ellis deserves further analysis.

Nanosized Alumina Particle and Proteasome Inhibitor Bortezomib Prevented inflammation and Osteolysis Induced by Titanium Particle via Autophagy and NF-κB Signaling.

Autophagy and NF-κB signaling are involving in the process of Particle Disease, which was caused by the particles released from friction interface of artificial joint, implant materials of particle reinforced composite, scaffolds for tissue engineering, or material for drug delivery. However, the biological interaction of different material particles and the mechanism of proteasome inhibitor, Bortezomib (BTZ), against Titanium (Ti) particle-induced Particle Disease remain unclear. In this study, we evaluated effect of nanosized Alumina (Al) particles and BTZ on reducing and treating the Ti particle-induced inflammatory reaction in MG-63 cells and mouse calvarial osteolysis model. We found that Al particles and BTZ could block apoptosis and NF- κB activation in osteoblasts in vitro and in a mouse model of calvarial resorption induced by Ti particles. We found that Al particles and BTZ attenuated the expression of inflammatory cytokines (IL-1ß, IL-6, TNF-α). And Al prevented the IL-1ß expression induced by Ti via attenuating the NF- κB activation ß-TRCP and reducing the expression of Casepase-3. Expressions of autophagy marker LC3 was activated in Ti group, and reduced by Al and/not BTZ. Furthermore, the expressions of OPG were also higher in these groups than the Ti treated group. Collectively, nanosized Al could prevent autophagy and reduce the apoptosis, inflammatory and osteolysis induced by Ti particles. Our data offered a basic data for implant design when it was inevitable to use Ti as biomaterials, considering the outstanding mechanical propertie of Ti. What's more, proteasome inhibitor BTZ could be a potential therapy for wear particle-induced inflammation and osteogenic activity via regulating the activity of NF- κB signaling pathway.

Chemical composition, antioxidant, antibacterial, and tyrosinase inhibition activity of extracts from Newhall navel orange (Citrus sinensis Osbeck cv. Newhall) peel.

BACKGROUND: Newhall navel orange (Citrus sinensis Osbeck cv. Newhall) is the major navel orange cultivar planted in China. Almost all Newhall navel orange peels produced in that country are currently discarded, which is not only harmful to the environment but also a waste of resources. It is therefore necessary to carry out research to explore the utilization potential of this resource. RESULTS: A 95% ethanol extract (EE) of Newhall orange peel was prepared and fractionated into three subextracts: petroleum ether extract (PEE), ethyl acetate extract (EtOAcE), and water extract (WE) by simple liquid / liquid extractions. These four extracts were then subjected to antioxidant, antibacterial, and tyrosinase inhibition assays. Interestingly, EtOAcE was significantly superior to all other three extracts, exhibiting the best biological effects. The total polyphenol content (TPC), total flavonoid content (TFC), and primary individual flavonoids of these four extracts were analyzed and compared. This was followed by principal component analysis (PCA) and the Pearson's correlation test. The result indicates that the primary bioactive compounds responsible for the biological effects of the EtOAcE are sinensetin, 4',5,6,7-tetramethoxyflavone, nobiletin, 3,3',4',5,6,7-hexamethoxyflavone, and narirutin. In view of its easy preparation and potent biological effects, EtOAcE might demonstrate excellent application potential in various industrial areas. CONCLUSIONS: This study successfully identified EtOAcE as a potent naturally occurring antioxidant, antibacterial, and tyrosinase inhibitory agent, which might add value to the utilization of Newhall navel orange peel in the food, cosmetic, and pharmaceutical industries. © 2020 Society of Chemical Industry.

Comparison of Two Numerical Parameters to Assess Detrusor Contractility in Prognosing Short-Term Outcome after Transurethral Resection of the Prostate.

OBJECTIVE: To investigate and compare the influence of two numerical detrusor contractility parameters, the bladder contractility index (BCI) and the maximum Watts factor (WFmax), on transurethral resection of the prostate (TURP) outcome. METHODS: A retrospective study was conducted on 236 patients who had undergone urodynamic assessment preoperatively and TURP for benign prostatic obstruction. They were evaluated by International Prostate Symptom Score (IPSS) and uroflowmetry preoperatively and 3 months postoperatively. Related criteria were established to determine the overall efficacy of TURP. Logistic regression analysis and receiver operating characteristic curves were made to investigate the influence of the BCI and WFmax on TURP efficacy. RESULTS: Among the 236 patients, 195 treatments were effective and 41 ineffective. Multivariate analysis showed that both the BCI (OR 1.038) and the WFmax (OR 1.291) could influence TURP efficacy. For predicting TURP efficacy, the optimal cut-off values of the BCI and WFmax were 98.7 and 10.27 W/m2, respectively. The AUC, sensitivity and specificity of the BCI were 0.722, 78.5% and 61.0%; those of the WFmax were 0.761, 73.9% and 73.2%, with no significant difference (p > 0.05). CONCLUSIONS: To some extent, the BCI and the WFmax can predict TURP efficacy equally well. A discrimination level of 10.27 W/m2 may be a threshold value for detrusor underactivity (DU); as regards the BCI, the current threshold value is appropriate to diagnose DU.

Effects of Wuzhi Capsules on Blood Concentration of Tacrolimus in Renal Transplant Recipients.

BACKGROUND Tacrolimus is a widely used immunosuppressant in renal transplant recipients. It was demonstrated in rats and healthy volunteers that Wuzhi capsules could inhibit metabolism and maintain blood concentration of tacrolimus. However, there are no clinical studies of Wuzhi capsules in renal transplant recipients. This research aimed to assess the effect of Wuzhi capsules on the blood concentration of tacrolimus in renal transplant recipients. MATERIAL AND METHODS A total of 158 Chinese renal transplant recipients receiving tacrolimus with or without Wuzhi capsules were included in this retrospective study. The cohort study included 126 recipients, with 86 recipients receiving Wuzhi capsules (WZCs) and the other 40 recipients not receiving WZCs. Another 32 recipients were involved in a self-control study. RESULTS Dose- and body weight-adjusted trough concentrations (C0/D/W) of tacrolimus in the WZC group were found to be significantly higher than that in the non-WZC group (P<0.05). The improvement of C0/D/W by administration of Wuzhi capsules was more significant in CYP3A5 expressers than in non-expressers following subgroup analysis. Furthermore, the WZC group had a remarkably higher proportion of subjects who reached target tacrolimus concentration than in the non-WZC group, both in CYP3A5 expressers (P=0.01) and non-expressers (P<0.001). Multiple linear regression analysis and self-control analysis confirmed the positive impact of Wuzhi capsules on tacrolimus concentration (P<0.001). CONCLUSIONS Wuzhi capsules can increase tacrolimus trough concentration without adverse effects on allograft function, especially in CYP3A5 expressers. Efficient and convenient immunosuppressive effects on renal transplant recipients can be achieved by treatment including administration of Wuzhi capsules.

Effects of Postharvest Time, Heat Treatment, pH and Filtration on the Limonin Content in Newhall Navel Orange (Citrus sinensis Osbeck cv. Newhall) Juice.

Delayed bitterness causes severe economic loss in citrus juice industry worldwide, which is mostly due to the formation of limonoid compounds, especially limonin, in juice. In this study, effects of postharvest time of fruits, heat treatment, pH and filtration of juice on limonin content in Newhall navel orange (Citrus sinensis Osbeck cv. Newhall) juice were investigated. Our research indicated for the first time that: (1) limonin content in juice would gradually increase to a maximal level and then remained almost constant thereafter as storage time going on, whereas the maximum constant value (MCV) of limonin content in juice significantly (p < 0.05) decreased with the increment of postharvest time of fruits being juiced; (2) heat treatment and acidification of juice only speeded up the formation of limonin to the maximal level while without changing the MCV of limonin content; (3) the juice after filtration exhibited much lower MCV of limonin content compared with the unfiltered one. These experimental observations might not only provide useful information for the development of new debitterness method for navel orange juice, but also strongly support the acid-promoted delayed bitterness mechanism, suggesting the formation of delayed bitterness might primary due to the acid-promoted rather than the enzyme-catalyzed lactonization of limonoate A-ring lactone (LARL) to produce limonin in juice of navel orange.

iTRAQ-based quantitative proteomic analysis reveals Bai-Hu-Tang enhances phagocytosis and cross-presentation against LPS fever in rabbit.

ETHNOPHARMACOLOGICAL RELEVANCE: Bai-Hu-Tang (BHT), a classical anti-febrile Chinese formula comprising of liquorice, anemarrhena rhizome, gypsum and rice, has been traditionally used to anti-febrile treatment and promote the production of body fluid to relieve thirst. In this paper, we aim to explore anti-febrile mechanism of BHT at protein level through analyzing alteration of differentially expressed proteins (DEPs) both lipopolysaccharide (LPS) fever syndrome and that was treated with BHT in rabbits. MATERIALS AND METHODS: Febrile model was induced by LPS injection (i.v.) in rabbits, and BHT (750mg dry extract/kg body weight) was gavaged to another group of LPS fever rabbits. After sacrifice of animals, total protein of liver tissue was isolated, and two-dimensional liquid chromatography (LC) - tandem mass spectrometry (MS) coupled with isobaric tags for relative and absolute quantification (iTRAQ) labeling analysis was employed to quantitatively identify differentially expressed proteins in two group animals, which were compared with control group. Then bioinformatic analysis of DEPs was conducted through hierarchical Clustering, Venn analysis, gene ontology (GO) annotation enrichment, and kyoto encyclopedia of genes and genomes (KEGG) pathways enrichment. RESULT: The results demonstrated there were 63 and 109 DEPs in LPS fever group and BHT-treated group, respectively. Enrichment analysis of GO annotations indicated that BHT mainly regulated expression of some extracellular structural proteins for response to stimulus and stress. KEGG analysis showed that ribosome and phagosome were the most significant pathways. Thereinto, several proteins in phagosome pathway were significantly up-regulated by BHT, including F-actin, coronin, Rac, and major histocompatibility complex class I (MHC I), which work in phagocytosis and cross-presentation CONCLUSION: BHT may contribute to pyrogen clearance by boosting antigenic phagocytosis, degradation, and cross presentation in the liver.

Analgesic and anti-inflammatory effects of hydroalcoholic extract isolated from Semen vaccariae.

Semen vaccariae, the seeds of Vaccaria segetalis (Neck.) Garcke, is usually used as an important medication for female mammary gland diseases; it has also been used to promote lactation for centuries in China. The purpose of this work was to evaluate the analgesic and anti-inflammatory effects of hydroalcoholic extract from semen vaccariae (HESV) with oral doses of 50, 100 and 200mg/kg•bw in mice and rats. We observed that the HESV could effectively inhibit acetic acid-induced abdominal contraction and could elevate the latency time to thermal stimuli in the hot-plate test in mice. In the xylene-induced ear-swelling test in mice, HESV could suppress the ear swelling. Additionally, HESV could significantly decrease the peritoneal capillary permeability and leukocyte infiltration in mice induced by the intraperitoneal injection of acetic acid. HESV also significantly reduced paw thickness 2-4 hours after the injection of carrageenan in the carrageenan-induced rat paw edema test. This study was the first to demonstrate that the oral administration of HESV might play an important role in the process of analgesia and anti-inflammation, supporting its use for female mammary gland diseases in traditional medicine.

Evaluation of bioaccumulation and toxic effects of copper on hepatocellular structure in mice.

The present study was to evaluate the hepatotoxicity effects in mice exposed to copper (Cu) used as dietary supplements for 95 days. Cu-treated mice showed increased body weight, and no toxic symptoms were observed at the beginning, but the tendency gradually changed with progress of experiment. In the liver, beneficial metals [Cu, iron (Fe), zinc (Zn), manganese (Mn), and molybdenum (Mo)] were analyzed by flame atomic absorption spectrometry. The content of Cu maintained at the same level during the experiments, but not resulting in the imbalance of Fe, Zn, Mn, and Mo being distributed. The activities of alkaline phosphatase (AKP) and super oxidation dismutase (SOD) showed significantly improvement during the first 30 days in Cu-supplemented group (P<0.01) but declined rapidly from 30th to 60th days, and later, they stabilized and were not statistically significant compared with control (P>0.05). No statistically significant correlation of ceruloplasmin (CPL) activity was appreciated during the experiment. The histopathological and ultrastructural abnormalities changes were observed in the liver of mice including vacuolar degeneration, necrosis, karyorrhexis, and endolysis. Many hepatocytes showed increased collagenic fibers, appearance of triglyceride droplets, and swollen mitochondria due to oral route of copper, which may lead to lipid peroxidation and free radicals. In conclusion, our study showed that exposure to copper influenced behavioral pattern and body weight, affected several enzymatic activities, and led to the physiological and considerable structural changes in the liver of mice. The public should pay more attention to avoid being exposed to copper.

Evaluation of potential effects of soil available phosphorus on soil arsenic availability and paddy rice inorganic arsenic content.

The transfer of arsenic from paddy field to rice is a major exposure route of the highly toxic element to humans. The aim of our study is to explore the effects of soil available phosphorus on As uptake by rice, and identify the effects of soil properties on arsenic transfer from soil to rice under actual field conditions. 56 pairs of topsoil and rice samples were collected. The relevant parameters in soil and the inorganic arsenic in rice grains were analyzed, and then all the results were treated by statistical methods. Results show that the main factors influencing the uptake by rice grain include soil pH and available phosphorus. The eventual impact of phosphorus is identified as the suppression of As uptake by rice grains. The competition for transporters from soil to roots between arsenic and phosphorus in rhizosphere soil has been a dominant feature.

Efficacy of herbal tincture as treatment option for retained placenta in dairy cows.

Retained placenta remains therapeutic challenge in cattle. Certain traditional medicines are believed to be able to alleviate retained placenta condition and improve overall fertility in cows. The aim of the present study was to evaluate the efficacy of an herbal tincture for treatment of retained placenta. The herbal tincture was extracted from a combination of Herba Leonuri, Angelicae Sinensis Radix, Flos Carthami, Myrrha and Rhizoma Cyperi by percolation with 70% ethanol to a concentration of 0.5g crude herb/ml. Cows diagnosed with retained placenta (n=48) were randomly divided into one of two treatment groups (A and B), with animals in group A (n=26) receiving herbal tincture orally, and cows in group B (n=22) receiving oxytetracycline infusion into the uterus. Eighty six cows with no clinically visible pathological conditions, given birth alone and with no retained placenta diagnosis were included into control group (C). Retained placenta was expelled within 72h following initial treatment in 19 cows in group A, yet no cows in group B were recorded to expel placenta in the same time. The median number of days to first service (70.0 vs. 102.5 days; P<0.05) and median number of days open (76.0 vs. 134.0 days; P<0.01) were lower in group A than in group B. Percentage of cows pregnant within 100 days postpartum was the highest for animals in group A compared to controls (61.5% vs. 39.5%, P<0.05), and for animals in group B (61.5% vs. 22.7%; P<0.01). Herbal tincture used in the present study might facilitate expulsion of retained placenta and improve subsequent fertility, thus could present effective treatment option for retained placenta in cows.

The administration of Sheng Hua Tang immediately after delivery to reduce the incidence of retained placenta in Holstein dairy cows.

Sheng Hua Tang, a classical herbal formula consisting of Radix Angelicae sinensis, Ligustici rhizoma, Semen persicae, Zingiberis rhizoma, and Radix glycyrrhizae, is known to be beneficial in alleviating postpartum diseases and facilitating a return to normal reproductive function. This study investigated whether the administration of Sheng Hua Tang within 2 to 4 hours after delivery was effective as a preventive treatment for reducing the risk of retained placenta in Holstein dairy cows. A total of 357 cows, each of which had delivered its calf spontaneously, were randomly allocated to one of two groups. In the treatment group, the cows (n = 175) received Sheng Hua Tang with an oral dose of 0.36 g crude herb per kg·body weight once daily for three consecutive days. The controls (n = 182) received no treatment. The placental retention proportion was 4.0% and 17.0% within 12 hours after delivery in the treated and control animals, respectively (P < 0.01). We found decreases in the calving-to-first-service interval (73.2 ± 25.1 vs. 81.9 ± 32.8 days; P < 0.01), calving-to-conception interval (93.4 ± 38.8 vs. 114.6 ± 42.9 days; P < 0.01), and service per conception (1.5 ± 0.8 vs. 1.9 ± 1.0 days; P < 0.01) in the treatment group compared with the control group. The first artificial insemination conception proportion was higher in the treatment group than in the control group (60.4% vs. 41.1%; P = 0.01). Moreover, the between-group difference in the proportion of cows that were pregnant within 180 days postpartum approached statistical significance (88.2% vs. 80.6%; P = 0.07). Sheng Hua Tang showed beneficial effects in reducing the incidence of retained placenta and improving subsequent reproductive performance in cows. This preventive treatment strategy would be effective in improving the management of puerperal health. The potential benefits of Sheng Hua Tang warrant further investigation to determine whether this preventive treatment strategy can be endorsed as a general preventive approach in postpartum cows.

Immunomodulatory activity in vitro and in vivo of polysaccharide from Potentilla anserina.

The polysaccharide (PAP) from Potentilla anserina was evaluated for modulating effects by using mouse peritoneal macrophage and the immunosuppressed-model cyclophosphamide-induced. Phagocytotic and mononuclear phagocytic system function assays showed that PAP stimulated the phagocytosis of phagocyte. Splenocyte proliferation assay showed that PAP acted the effect combining Con A or LPS in splenocyte proliferation. The parameters detected showed that PAP increased thymus and spleen indices, the levels of LDH and ACP in the spleen, and IL-10 and IFN-γ in serum in immunosuppressed mice. The results suggest that PAP is involved in immunomodulatory effects leading to the exploration for PAP as a potential immunostimulant.

[Dynamic study on effect of Chinese medicine compound Yiliuyin on angiogenesis in transplanted S180 tumor of mouse].

OBJECTIVE: To investigate the dynamic changes in angiogenesis within the tumor tissue of mice bearing S180 tumor at different day-points of oral administration with a Chinese medicine compound "Yiliuyin" (YLY) and to explore the anti-tumor mechanisms of YLY in vivo. METHOD: Fifty-six BALB/c mice were divided into YLY group and control group (28 mice/group) and each group was divided into four subgroups (7 mice/subgroup), randomly. After 24 hrs of inoculation with S180 tumor cells subcutaneously in the right axilla, YLY in the mice of YLY group and equal volume of cold boiled-water in the mice of control group were administered orally twice every day, 0.5 mL each time. The mice of one subgroup from the two groups apiece were killed at 10, 20, 30 th and 40 th day-point of oral administration, respectively. The tumors were isolated and were made into paraffin embedded sections. The dynamic changes of the angiogenesis (CD34 staining), vascular endothelial growth factor (VEGF), vascular endothelial growth factor receptor-2 (VEGFR-2) and endostatin (ES) in tumor tissue were detected by immunohistochemistry staining, and the results were shown as PED (positive enzyme dot). RESULT: YLY could remarkably decrease the angiogenesis within tumor tissues. The PED of CD34 in control group at 10, 20, 30 th and 40 th day-point was 392.86+/-42.01, 481.49+/-58.34, 386.31+/-54.91 and 376.69+/-28.71, and that in YLY group was 334.46+/-33.38, 289.34+/-39.63, 257.09+/-40.00 and 246.57+/-36.78, respectively. The PED of CD34 in YLY group at each day-point was lower than that in control group (P<0.05, P<0.01, P<0.01 and P<0.01, respectively). The PED of VEGF in control group at 10, 20, 30 th and 40 th day-point was 852.63+/-81.65, 1168.40+/-96.69, 1292.60+/-147.54 and 1124.74+/-139.64, and that inYLY group was 718.40+/-94.94, 866.54+/-72.40, 859.31+/-74.02 and 753.34+/-72.95, respectively. The PED of VEGF in YLY group at each day-point was lower than that in control group (P <0.05, P <0.01, P <0.01 and P <0.01, respectively). The PED of VEGFR-2 in control group at 10th, 20th, 30th and 40th day-point was 618.63+/-59.08, 750.09+/-56.72, 684.91+/-72.86 and 644.06+/-60.25, and that in YLY group was 523.91+/-64.66, 449.03+/-46.85, 400.06+/-60.12 and 339.89+/-45.39, respectively. The PED of VEGFR-2 in YLY group at each day-point was lower than that in control group (P <0.05, P <0.01, P <0.01 and P <0.01, respectively). The PED of ES in control group at 10th, 20th, 30th and 40th day-point was 250.26+/-36.27, 298.60+/-44.41, 450.86+/-38.95 and 398.43+/-34.19, and that in YLY group was 249.57+/-40.23, 350.03+/-40.92, 499.40+/-40.29 and 497.94+/-42.76, respectively. There was no difference between the two groups at 10th day-point.The PED of ES in YLY group was higher than that in control group at 20, 30, 40 th day-point (P <0.05, P <0.01 and P <0.01, respectively) . CONCLUSION: YLY could exert the anti- tumor role by down-regulating the expression of VEGF and VEGFR-2, up-regulating the expression of ES and inhibiting the angiogenesis within tumor tissue.

Discovery and X-ray crystallographic analysis of a spiropiperidine iminohydantoin inhibitor of beta-secretase.

A high-throughput screen at 100 microM inhibitor concentration for the BACE-1 enzyme revealed a novel spiropiperidine iminohydantoin aspartyl protease inhibitor template. An X-ray cocrystal structure with BACE-1 revealed a novel mode of binding whereby the inhibitor interacts with the catalytic aspartates via bridging water molecules. Using the crystal structure as a guide, potent compounds with good brain penetration were designed.

Discovery of 1,4-substituted piperidines as potent and selective inhibitors of T-type calcium channels.

The discovery of a novel series of potent and selective T-type calcium channel antagonists is reported. Initial optimization of high-throughput screening leads afforded a 1,4-substituted piperidine amide 6 with good potency and limited selectivity over hERG and L-type channels and other off-target activities. Further SAR on reducing the basicity of the piperidine and introducing polarity led to the discovery of 3-axial fluoropiperidine 30 with a significantly improved selectivity profile. Compound 30 showed good oral bioavailability and brain penetration across species. In a rat genetic model of absence epilepsy, compound 30 demonstrated a robust reduction in the number and duration of seizures at 33 nM plasma concentration, with no cardiovascular effects at up to 5.6 microM. Compound 30 also showed good efficacy in rodent models of essential tremor and Parkinson's disease. Compound 30 thus demonstrates a wide margin between CNS and peripheral effects and is a useful tool for probing the effects of T-type calcium channel inhibition.