Evaluation of the anti-proliferative properties of selected psoriasis-treating Chinese medicines on cultured HaCaT cells

Psoriasis is a chronic inflammatory skin disorder, which affects approximately 2-3% of the population worldwide. The current conventional therapy cannot offer satisfactory clinical results for most of the patients, largely due to the fact that many anti-psoriatic drugs have serious side effects and psoriasis is prone to developing drug resistance after long term exposure. Traditionally, Chinese herbal medicine has been extensively used to treat psoriasis and produced promising clinical results; however, its underlying mechanisms of action have not been systematically investigated. The aim of this study was to investigate those Chinese medicinal materials, which are commonly prescribed in Chinese medicine practice for psoriasis, for their anti-proliferative effects on HaCaT cells in vitro. Sixty Chinese medicinal materials were selected and extracted with 80% aqueous ethanol. The dry extracts were evaluated for their anti-proliferative activities by microplate SRB and MTT assays. Three Chinese medicinal materials i.e. the root of Rubia cordifolia L. (Rubiaceae), Realgar and the rhizome of Coptis chinensis Franch. (Ranunculaceae) were found to have significant anti-proliferative effects, with IC(50) being 1.4, 6.6 and 23.4 microg/ml, respectively as measured by MTT assay. While Realgar was also able to modestly inhibit the growth of Hs-68 cells in vitro, Rubia cordifolia and Coptis chinensis did not exert cytotoxicity to this human fibroblast cell line.     

三磷酸腺苷氯化镁对肺癌细胞(A549)端粒酶活性的影响

目的:探讨三磷酸腺苷氯化镁(ATP-MgCl2)对肺腺癌细胞的影响及其作用机理.方法:采用连续细胞记数方法观测ATP-MgCl2对A549肺腺癌细胞生长的抑制情况,应用端粒重复序列扩增法(TRAP)观察ATP-MgCl2对癌细胞端粒酶活性的影响,并与ATP作对比.结果:ATP-MgCl2显著抑制癌细胞生长(P＜0.01),且使大量癌细胞死亡,而加药后前两天ATP-MgCl2的抑制、杀伤癌细胞程度较ATP明显(P＜0.05).ATP-MgCl2组和ATP组的端粒酶阳性表达率明显低于对照组(P＜0.01),而前两者间无明显差异(P＜0.05).结论:ATP-MgCl2对肺腺癌细胞有抑制、杀伤(细胞毒)作用.其抗癌作用机理与ATP相似,与抑制癌细胞端粒酶活性有关.MgCl2与ATP结合形成一种活性复合物,增强了ATP的抗癌作用,ATP-MgCl2具有较好的临床应用价值.     

Self-reported yeast symptoms compared with clinical wet mount analysis and vaginal yeast culture in a specialty clinic setting

OBJECTIVE: To determine the validity of patients' self-reported symptoms of vulvovaginal candidiasis and the accuracy of clinical wet mount examinations compared with vulvovaginal yeast culture results in a specialty clinic. STUDY DESIGN: A retrospective chart review of new patients seen at the Saint Louis University Vulvar and Vaginal Disease Clinic from January 2005 to March 2006 was performed. Patients' age, medication use, symptom scores on a rating scale for vaginal/vulvar pain, burning, itching, dyspareunia and wet mount analyses were compared with yeast culture results. RESULTS: Of 153 patients, 40 had positive yeast cultures (prevalence rate 26.1%). Compared with yeast cultures, self-reported symptom scores >4 resulted in high sensitivity (90%) and low specificity (7%). Positive wet mount result showed low sensitivity (18%) and high specificity (99%). Patient symptom scores were a poor predictor of yeast infections based on yeast culture results. No correlation was found among wet mount, self-reported symptoms and yeast culture results. No significant difference between age or symptom scores to culture result was found. CONCLUSION: Wet mount analysis for recurrent or persistent patient symptoms should be reevaluated. Self-reported symptoms are not reliable for diagnosis. Wet mount analysis resulted in low sensitivity. Yeast cultures should be considered the gold standard for identification of vulvovaginal candidiasis in persistent or recurrent cases.     

Differential flow cytometric detection of intracellular cytokines in peripheral and peritoneal mononuclear cells of women with endometriosis

OBJECTIVE: The pathogenesis of endometriosis is related to functional changes in CD3+ and CD14+ cells observed both at the local and systemic level. Here we investigated whether, and if so, how the body compartment influences cytokine expression in stimulated peritoneal and peripheral CD3+ and CD14+ cells of women with endometriosis. STUDY DESIGN: Isolated peripheral blood (PB) and peritoneal fluid (PF) mononuclear cells from women with endometriosis were cultured under non-adherent conditions and stimulated with PMA and ionomycin for 6h to induce intracellular cytokine synthesis of TNF-alpha, IFN-gamma, and IL-8 by CD3+ cells or with LPS for 9h to produce TNF-alpha, IL-6, IL-10, MCP-1, and IL-8 by CD14+ cells. RESULTS: The percentages of positive CD3+ cells stained for TNF-alpha and IFN-gamma were significantly higher and those stained for IL-8 were significantly lower in PF compared with PB, this being independent of the stage of endometriosis. In contrast, the percentages of CD14+ cells producing TNF-alpha, IL-6, IL-10, MCP-1, and IL-8 were significantly higher in PB than PF of women with endometriosis. CONCLUSIONS: Monocytes/macrophages and lymphocytes derived from the peripheral and peritoneal compartments of women with endometriosis differentially respond to stimulated cytokine synthesis induction. However, it is difficult to state whether the observed phenomenon is more related to body compartment influence per se or to the presence of endometriosis.     

阿托伐他汀联合雄激素阻断对晚期前列腺癌术后患者血清PSA、VEGF、MMP-9及血脂水平的影响

目的观察阿托伐他汀联合雄激素阻断治疗对晚期前列腺癌患者血清前列腺特异性抗原(PSA)、血管内皮生长因子(VEGF)、基质金属蛋白酶-9(MMP-9)及血脂水平的影响。方法选取2016年1-12月海南医学院第二附属医院泌尿外科一区治疗晚期前列腺癌患者60例,均先行经尿道前列腺电切术(TURP),根据随机数字表法分为2组,每组30例。对照组术后以间断雄激素阻断治疗,观察组以阿托伐他汀联合间断雄激素阻断治疗。比较2组治疗前后血清PSA、VEGF、MMP-9、血脂水平。并随访3年,观察2组患者生存率及不良反应发生率。结果治疗后,观察组患者PSA、VEGF和MMP-9水平低于对照组(t/P=8.328/0.000、2.531/0.014、9.680/0.000);TG、TC、LDLC和HDL-C明显优于对照组(t/P=5.350/0.000、9.348/0.000、21.601/0.000、2.402/0.000)胃肠道反应、性欲减退、肝功能异常、勃起障碍及骨质疏松症等不良反应发生情况与对照组患者比较,差异无统计学意义(P>0.05);1、2、3年生存率和不良反应发生率优于对照组(χ^2/P=4.286/0.038、4.812/0.028、4.800/0.028、4.800/0.028、4.800/0.028、4.893/0.032)。结论阿托伐他汀联合雄激素阻断治疗方案可以有效降低晚期前列腺癌患者血清PSA、VEGF和MMP-9水平,改善血脂水平,提高生存率,安全性较好。     

Berberine improves intralipid-induced insulin resistance in murine

Insulin resistance (IR) is a major metabolic risk factor even before the onset of hyperglycemia. Recently, berberine (BBR) is found to improve hyperglycemia and IR. In this study, we investigated whether BBR could improve IR independent of hyperglycemia. Acute insulin-resistant state was induced in rats by systemic infusion of intralipid (6.6%). BBR was administered via different delivery routes before or after the beginning of a 2-h euglycemic-hyperinsulinemic clamp. At the end of experiment, rats were sacrificed, gastrocnemius muscle was collected for detecting mitochondrial swelling, phosphorylation of Akt and AMPK, as well as the mitochondrial permeability regulator cyclophilin D (CypD) protein expression. We showed that BBR administration markedly ameliorated intralipid-induced IR without affecting blood glucose, which was accompanied by alleviated mitochondrial swelling in skeletal muscle. We used human skeletal muscle cells (HSMCs), AML12 hepatocytes, human umbilical vein endothelial cells, and CypD knockout mice to investigate metabolic and molecular alternations. In either HSMCs or AML12 hepatocytes, BBR (5 μM) abolished palmitate acid (PA)-induced increase of CypD protein levels. In CypD-deficient mice, intralipid-induced IR was greatly attenuated and the beneficial effect of BBR was diminished. Furthermore, we demonstrated that the inhibitory effect of BBR on intralipid-induced IR was mainly mediated by skeletal muscle, but not by intestine, liver, or microvasculature; BBR administration suppressed intralipid-induced upregulation of CypD expression in skeletal muscle. These results suggest that BBR alleviates intralipid-induced IR, which is related to the inhibition of CypD protein expression in skeletal muscle.     

Effects of Chronic Exposure to Diets Containing Moldy Corn or Moldy Wheat Bran on Growth Performance,Ovarian Follicular Pool,and Oxidative Status of Gilts

Background: We investigated the effect of replacing normal corn (NC) or normal wheat bran (NW) with moldy corn (MC) or moldy wheat bran (MW) on growth, ovarian follicular reserves, and oxidative status. Methods: Sixty-three Landrace × Yorkshire gilts were assigned to seven diets formulated by using MC to replace 0% (control), 25% (25% MC), 50% (50% MC), 75% (75% MC), and 100% NC (100% MC), MW to replace 100% NW (100% MW), and MC and MW to replace 100% NC and 100% NW (100% MC + MW), from postnatal day 110 to day 19 of the second estrous cycle. Results: Feeding the gilts with MC or MW induced a lower average daily gain at days 29–56 of the experiment. Age at puberty remained unchanged, but MC inclusion resulted in a linear decrease in antral follicles with diameter >3.0 mm, and control gilts had a 12.7 more large antral follicles than gilts in the 100% MC + MW treatment. MC inclusion linearly decreased the numbers of primordial follicles, growing follicles, and corpora lutea, associated with a lower anti-Müllerian hormone level in serum and 17β-estradiol level in follicular fluid. MC inclusion decreased the serum concentrations of insulin-like growth factor 1 and its mRNA levels in the liver, combined with higher malondialdehyde concentration and lower total superoxide dismutase activities in serum and liver. Conclusion: Chronic exposure to MC-containing diets caused the loss of follicles, even if levels of deoxynivalenol, zearalenone, and aflatoxin B1 were below the levels allowed by China and Europe standards.     

Discovery of a highly specific 18F-labeled PET ligand for phosphodiesterase 10A enabled by novel spirocyclic iodonium ylide radiofluorination

As a member of cyclic nucleotide phosphodiesterase (PDE) enzyme family, PDE10A is in charge of the degradation of cyclic adenosine (cAMP) and guanosine monophosphates (cGMP). While PDE10A is primarily expressed in the medium spiny neurons of the striatum, it has been implicated in a variety of neurological disorders. Indeed, inhibition of PDE10A has proven to be of potential use for the treatment of central nervous system (CNS) pathologies caused by dysfunction of the basal ganglia–of which the striatum constitutes the largest component. A PDE10A-targeted positron emission tomography (PET) radioligand would enable a better assessment of the pathophysiologic role of PDE10A, as well as confirm the relationship between target occupancy and administrated dose of a given drug candidate, thus accelerating the development of effective PDE10A inhibitors. In this study, we designed and synthesized a novel ¹⁸F-aryl PDE10A PET radioligand, codenamed [¹⁸F]P10A-1910 ([¹⁸F]9), in high radiochemical yield and molar activity via spirocyclic iodonium ylide-mediated radiofluorination. [¹⁸F]9 possessed good in vitro binding affinity (IC₅₀ = 2.1 nmol/L) and selectivity towards PDE10A. Further, [¹⁸F]9 exhibited reasonable lipophilicity (logD = 3.50) and brain permeability (P_app > 10 × 10⁻⁶ cm/s in MDCK-MDR1 cells). PET imaging studies of [¹⁸F]9 revealed high striatal uptake and excellent in vivo specificity with reversible tracer kinetics. Preclinical studies in rodents revealed an improved plasma and brain stability of [¹⁸F]9 when compared to the current reference standard for PDE10A-targeted PET, [¹⁸F]MNI659. Further, dose–response experiments with a series of escalating doses of PDE10A inhibitor 1 in rhesus monkey brains confirmed the utility of [¹⁸F]9 for evaluating target occupancy in vivo in higher species. In conclusion, our results indicated that [¹⁸F]9 is a promising PDE10A PET radioligand for clinical translation.KEY WORDS: Phosphodiesterase 10A, PET radioligand, ¹⁸F, Spirocyclic iodonium ylide, Nonhuman primate, Target occupancy