Synthesis of bilocularin A carbamate derivatives and their evaluation as leucine transport inhibitors in prostate cancer cells.

Large-scale extraction of the leaves of the Australian rainforest tree Maytenus bilocularis followed by extensive purification studies afforded the targeted and abundant dihydro-ß-agarofuran, bilocularin A, in sufficient quantities (>500 mg) for detailed semi-synthetic chemistry. Eight bilocularin A carbamate analogues were synthesised using a series of commercially available isocyanate reagents in high purity (>95%) and variable yields (9-91%). All previously undescribed analogues were spectroscopically characterised using NMR, UV, IR and MS data. One compound afforded crystalline material and subsequent single crystal X-ray analysis (Cu-Kα) confirmed the chemical structure along with the absolute configuration. All compounds were evaluated for anti-proliferative activity against the human prostate cancer cell line LNCaP; none of the compounds showed significant (>50%) growth inhibition at 20 µM. Compounds were also tested for their ability to inhibit leucine transport in LNCaP cells, and two analogues showed moderate activity with IC50 values of 8.9 and 8.5 µM. This is the first reported synthesis of dihydro-ß-agarofuran carbamate derivatives.

[Moxibustion improved gastric ulcer by reducing contents of corticotrophin-releasing hormone and adrenocorticotropic hormone in serum and hypothalamus-pituitary tissues in rats with stress-induced gastric ulcer].

OBJECTIVE: To observe the effect of moxibustion of "Zhongwan" (CV12) and "Zusanli" (ST36) on histopathological changes of the gastric mucosa and contents of corticotropin-releasing hormone(CRH) and adrenocorticotrophic hormone(ACTH) in the serum, hypothalamus and pituitary tissues in rats with stress-induced gastric ulcer(SGU), so as to reveal its mechanisms underlying improvement of SGU. METHODS: A total of 28 male SD rats were randomly divided into blank control, model, moxibustion, and medication groups, with 7 rats in each group. The SGU model was established by water immersion restraint stress for 3 h. Moxibustion was applied to "Zhongwan"(CV12) and bilateral "Zusanli" (ST36) for 20 min, once a day for 5 days, and rats of the medication group were treated by gavage of Omeprazole enteric-coated tablets (0.2 mg/kg) once a day for 5 days. The gastric mucosal damage index (ulcer index, UI) was measured to assess the injury severity according to Guth's me-thods. Histopathological changes of the gastric mucosa were determined by H．E. staining. The contents of CRH in serum and hypothalamus and ACTH in serum and pituitary gland tissue were assayed by using ELISA. RESULTS: Outcomes of H．E. staining showed gastric mucosal epithelia defect, disordered arrangement of glands, obvious mucosal hyperemia and edema, exudation of a large number of red blood cells, swelling of mucosal cells with necrosis of nuclei in the model group. These situations were relatively milder in the moxibustion and medication groups. After modeling, the UI, and the contents of CRH in the serum and hypotha-lamus, and ACTH in the serum and pituitary tissue were significantly increased in comparison with the blank control group (P<0.01). Following the intervention, the UI, and contents of CRH in the serum and hypothalamus, and ACTH in the serum and pituitary were all down-regulated in both medication and moxibustion groups relevant to the model group (P<0.05, P<0.01). The therapeutic effect of moxibustion was notably superior to that of medication in down-regulating serum CRH and ACTH (P<0.05). No significant differences were found between the medication and moxibustin interventions in lowering the UI, hypothalamic CRH and pituitary ACTH levels (P>0.05)．. CONCLUSION: Moxibustion can relieve gastric mucosal injury induced by stress in water immersion restraint stress rats, which may be associated with its effects in down-regulating the levels of CRH and ACTH in se-rum, hypothalamus and pituitary tissues (inhibition of activities of hypothalamic-pituitary-adrenocortical axis)．.

[Effect of Electroacupuncture on Cerebro-cortex Caspase-3 Expression and Blood Lipid Levels in Hyperlipemia Rats with Cerebral Ischemia].

OBJECTIVE: To observe the effect of electroacupuncture (EA) stimulation of "Fenglong" (ST 40), "Sanyinjiao" (SP 6) plus manual acupuncture (MA) stimulation of "Shuigou" (GV 26) and "Baihui" (GV 20) on Caspase-3 protein expression in the cerebral cortex of rats with hyperlipemia and cerebral ischemia(HL-CI),so as to reveal its mechanisms underlying improvement of HL-CI. METHODS: Forty-five rats were randomly divided into normal control,sham operation,model,EA group I(EA+MA was given for 14 days, i.e., 7 days before CI, and 7 days more after HL-CI)and EA group Ⅱ (EA+MA was given for only 7 days after HL-CI),with 9 rats being in each group. The HL-CI model was established by feeding the animals with high fat forage for 6 weeks and then making an occlusion of the unilateral middle cerebral artery by regional application of quantitative paper adsorbing 50% FeCl3 solution (10 µL). Rats of the sham operation group were treated with the same procedures only without application of FeCl3 solution. For rats of the EA group I,EA (1-3 mA, 2 Hz/100 Hz) was applied to bilateral acupoints SP 6 and ST 40 (for 20 min),and MA stimulation applied to GV 26 and GV 20. EA was conducted once daily for 7 days after 6 weeks' high fat fo-rage feeding, and EA+MA intervention was conducted once daily for 7 days after CI modeling. For rats in the EA group Ⅱ, EA+MA was applied to the same 4 acupoints once a day for 7 days only after CI modeling. The neurological impairment was assessed by Zea Longa's scoring. The blood sample was taken from the abdominal aorta for measuring the contents of serum cholesterol (CHO),triglyceride (TG) and high-density lipoprotein cholesterol (HDL-C) and low-density lipoprotein cholesterol (LDL-C). Pathological changes of the cerebral cortex were observed after H.E. staining, and the expression of cerebro-cortex Caspase-3 was analyzed by immunohistochemistry. RESULTS: Following modeling,the neurological score,CHO, TG and LDL-C contents, and the number of Caspase-3 positive cells as well as Caspase-3 immunoactivity level were significantly increased in the model group(P<0.05), while serum HDL-C level was obviously decreased(P<0.05). After the treatment,the increased neurological score, CHO, TG and LDL-C contents, and the number of Caspase-3 positive cells and Caspase-3 immunoactivity level were considerably decreased in the EA group I and Ⅱ(P<0.05)while the decreased HDL-C level was notably increased relevant to the model group(P<0.05). The effects of the EA group I were notably superior to those of EA group Ⅱ in down-regulating the neurological score,CHO, TG and LDL-C levels and the expression of Caspase-3 protein(P<0.05). No significant differences were found between the normal control and sham operation groups in the neurological scores 20 min and 7 days after modeling and Caspase-3 expression levels (P>0.05). H.E. staining showed a reduction of the apoptotic cells and inflammatory cells in both EA group I and Ⅱ. CONCLUSIONS: Both EA and EA+MA interventions can improve neurological function in HL-CI rats,which may be related to their effects in adjusting the levels of serum lipids and down-regulating the expression of cell apoptosis-related Caspase-3 protein in the ischemic cortex. Moreover, the cerebral ischemia injury may be lightened by EA-lowering hyperlipemia first.

[Systematic Evaluation and Meta-analysis on Acupuncture for Peptic Ulcer].

OBJECTIVE: To systematically evaluate the clinical efficacy of acupuncture for peptic ulcer. METHODS: Randomized controlled trials of acupuncture for peptic ulcer were searched from China National Knowledge Infrastructure (CNKI), WanFang Database, Chinese Scientific and Technological Journals (VIP), China Biomedicine (CBM), PubMed and the Cochrane Library from the establishment time of databases to September, 2016. Data extraction and quality evaluation were implemented for the literature which met the inclusive criteria. The RevMan 5.3 software was used to make Meta-analysis. RESULTS: Sixteen papers including 1 570 patients of peptic ulcer were included. The results of Meta-analysis showed that there was no statistical significance between acupuncture and western medicine in the effective rate, the healing rate of ulcer area and the HP negative rate (all P>0.05); the recurrence rate of acupuncture was significantly lower than that of western medicine[RR=0.35, 95%CI (0.14, 0.84), P<0.05]. Acupuncture plus western medicine was significantly different from simple western medicine in the effective rate, the healing rate of ulcer area and the recurrence rate[RR=1.20,95% CI (1.04, 1.38), P=0.01; RR=1.29, 95% CI (1.06, 1.58), P=0.01;RR=0.27, 95% CI (0.16, 0.45), P<0.00001]. The analysis of evidence grade (GRADE) pre-sented that the healing rate of ulcer area and the HP negative rate of acupuncture were "low grade", and others were "extremely low grade". CONCLUSIONS: Acupuncture combined with western medicine has some advantages for peptic ulcer compared with the conventional western medicine, which needs further confirmation due to the lower evidence grade. Larger samples, randomized controlled trials with high quality are highly recommended.

[Effects of Direct Moxibustion of "Ganshu"(BL 18) on the Contents of T cells in Peripheral Blood in Rats with Precancerous Lesion of Primary Hepatocellular Carcinoma].

OBJECTIVE: To observe the effect of moxibustion stimulation of "Ganshu"(BL 18) region on contents of T cells in the peripheral blood in rats with Diethylnitrosamine (DEN)-induced hepatocellular carcinoma (HCC), so as to explore its effective in improving immunoregulatory function. METHODS: Seventy male Wistar rats were randomly divided into control group (n=10), model group(n=15), direct moxibustion-15 s group(n=15), direct moxibustion-30 s group (n=15) and ginger-separated moxibustion group(n=15). The primary HCC precancerous lesion model was established by intraperitoneal injection of DEN (50 mg/kg), once every 3 days for 10 weeks. Moxibustion was applied to bilateral "Ganshu"(BL 18) for about 15 min (3 moxa-cones), or 30 min (six moxa-cones), with or without ginger-slice separation, once every other day for 10 consecutive weeks. The contents of T cells of CD 3+, CD 4+, CD 8+ and CD 4+/CD 8+ ratio in the peripheral blood were detected with Flow Cytometey(FCM), pathological changes of liver were observed by light microscope after hematoxylin-eosin(HE) stain. RESULTS: Compared to the control group, the contents of blood CD 3+ and CD 4+ T cells and ratio of CD 4+/CD 8+ were significantly down-regulated, while that of CD 8+T cells was obviously increased in the model group(P<0.05,P<0.01). After moxibustion intervention, the decreased CD 3+ and CD 4+T cells and CD 4+/CD 8+ levels and the increased CD 8+T cell contents were reversed in all the 3 moxibustion groups (P<0.05, P<0.01), except CD 3+ in the ginger-separated moxibustion group (P>0.05). There were no significant differences among the three moxibustion groups in the CD 3+, CD 4+, CD 8+ and CD 4+/CD 8+ levels (P>0.05). In addition, the pathological changes of liver tissue as central vein deviation or absence, disordered arrangement of hepatic cords, narrowing of the hepatic sinusoid, hyperplasia of collagen fibers, formation of tuberosis, unevenness of liver cells with nuclear anachromasis and higher heteromorphism, and macronucleus oncocytes in primary HCC rats were not observed or milder after moxibustion intervention. CONCLUSIONS: Direct moxibustion and ginger-separated moxibustion can improve pathological changes of hepatic cells in rats with HCC, which may be associated with its actions in raising blood CD 3+ and CD 4+ T cell contents and reducing CD 8+ T levels to improve immune function.