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.     

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     

Tei指数对高血压不同构型组心功能临床研究

目的评价心功能指标Tei指数在高血压患者各个不同构型组中的应用价值。方法对高血压组82例和正常对照组19例进行常规心脏超声、常规心脏收缩功能、心脏舒张功能指标及Tei指数测量。将原发性高血压划分为正常左室构型组Ⅰ、向心性重构组Ⅱ、向心性肥厚组Ⅲ、离心性肥厚组Ⅳ。结果(1)舒张功能:E/A值在Ⅰ、Ⅱ、Ⅲ组依次降低,Ⅳ组E/A反而增高,E/A>1。IVRT、EDT值高血压组各组间差异无显著意义。(2)收缩功能:EF、FS值在Ⅳ组才出现降低,SV和CO在Ⅰ和Ⅲ中无明显变化,Ⅱ减少,Ⅳ稍增加。(3)Tei指数高血压组较对照组皆增高,差异有显著意义。Ⅰ、Ⅱ、Ⅲ、Ⅳ组的Tei指数依次增高,除Ⅰ、Ⅱ组外,余各组之间差异有显著意义,Ⅳ组Tei值最高。结论Tei指数能早期发现反映整体功能的降低。这弥补了心脏舒张功能指标和收缩功能对高血压患者心功能评价的一些不足。     

Optimization of photo-biomodulation therapy for wound healing of diabetic foot ulcers in vitro and in vivo

Unclear optical parameters make photo-biomodulation (PBM) difficult to implement in diabetic foot ulcer (DFU) clinically. Here, 12 wavelengths (400–900 nm) were used to conduct PBM to heal DFU wounds in vitro and in vivo. PBM at 10 mW/cm² and 0.5–4 J/cm² with all 12 wavelengths promoted proliferation of diabetic wound cells. In a mimic DFU (mDFU) rat model, PBM (425, 630, 730, and 850 nm, and a combination light strategy) promoted mDFU healing. The positive cell proliferation, re-epithelialization, angiogenesis, collagen synthesis, and inflammation were possible mechanisms. The combination strategy had the best effect, which can be applied clinically.     

Gasdermin E-derived caspase-3 inhibitors effectively protect mice from acute hepatic failure

Programmed cell death(PCD),including apoptosis,apoptotic necrosis,and pyroptosis,is involved in various organ dysfunction syndromes.Recent studies have revealed that a substrate of caspase-3,gasdermin E(GSDME),functions as an effector for pyroptosis;however,few inhibitors have been reported to prevent pyroptosis mediated by GSDME.Here,we developed a class of GSDME-derived inhibitors containing the core structure of DMPD or DMLD.Ac-DMPD-CMK and Ac-DMLD-CMK could directly bind to the catalytic domains of caspase-3 and specifically inhibit caspase-3 activity,exhibiting a lower IC50 than that of Z-DEVD-FMK.Functionally,Ac-DMPD/DMLD-CMK substantially inhibited both GSDME and PARP cleavage by caspase-3,preventing apoptotic and pyroptotic events in hepatocytes and macrophages.Furthermore,in a mouse model of bile duct ligation that mimics intrahepatic cholestasis-related acute hepatic failure,Ac-DMPD/DMLD-CMK significantly alleviated liver injury.Together,this study not only identified two specific inhibitors of caspase-3 for investigating PCD but also,more importantly,shed light on novel lead compounds for treating liver failure and organ dysfunctions caused by PCD.