Stereoselective binding of doxazosin enantiomers to plasma proteins from rats,dogs and humans in vitro

Aim： （＋）Doxazosin is a long-lasting inhibitor of a1-adrenoceptors that is widely used to treat benign prostatic hyperplasia and lower urinary tract symptoms. In this study we investigated the stereoselective binding of doxazosin enantiomers to the plasma proteins of rats, dogs and humans in vitro. Methods： Human, dog and rat plasma were prepared. Equilibrium dialysis was used to determine the plasma protein binding of each enantiomer in vitro. Chiral HPLC with fluorescence detection was used to measure the drug concentrations on each side of the dialysis membrane bag. Results： Both the enantiomers were highly bound to the plasma proteins of rats, dogs and humans [（-）doxazosin： 89.4%-94.3%; （＋）doxazosin： 90.9%-95.4%]. （＋）Doxazosin exhibited significantly higher protein binding capacities than （-）doxazosin in all the three species, and the difference in the bound concentration （Cb） between the two enantiomers was enhanced as their concentrations were increased. Although the percentage of the plasma protein binding in the dog plasma was significantly lower than that in the human plasma at 400 and 800 ng/mL, the corrected percentage of plasma protein binding was dog〉human〉rat. Conclusion： （-）Doxazosin and （＋）doxazosin show stereoselective plasma protein binding with a significant species difference among rats, dogs and humans.     

Prenatal nicotine exposure enhances the susceptibility to metabolic syndrome in adult offspring rats fed high-fat diet via alteration of HPA axis-associated neuroendocrine metabolic programming

Aim： Prenatal nicotine exposure （PNE） alters the hypothalamic-pituitary-adrenocortical （HPA） axis-associated neuroendocrine metabolic programming in intrauterine growth retardation offspring rats. In this study we aimed to clarify the susceptibility to metabolic diseases of PNE offspring rats fed a high-fat diet. Methods： Maternal Wistar rats were injected with nicotine （1.0 mg/kg, sc） twice per day from gestational day 11 until full-term delivery, and all pups were fed a high-fat diet after weaning and exposed to unpredictable chronic stress （UCS） during postnatal weeks 18-20. Blood samples were collected before and after chronic stress, and serum ACTH, corticosterone, glucose, insulin, total cholesterol, triglyceride and free fatty acids levels were measured. The hypothalamus, pituitary gland and liver were dissected for histological studies. Results： UCS significantly increased the serum ACTH, corticosterone and insulin levels as well as the insulin resistant index without changing the serum glucose, total cholesterol, triglyceride and free fatty acids levels in adult offspring rats without PNE. The body weight of PNE offspring rats presented a typical ＂catch-up＂ growth pattern. PNE not only aggravated the UCS-induced changes in the HPA axis programmed alteration （caused further increases in the serum ACTH and corticosterone levels）, but also significantly changed the glucose and lipid metabolism after UCS （caused further increases in the serum glucose level and insulin resistant index, and decrease in the serum free fatty acids）. The effects of PNE on the above indexes after UCS showed gender differences. Pathological studies revealed that PNE led to plenty of lipid droplets in multiple organs. Conclusion： PNE enhances not only the HPA axis, but also the susceptibility to metabolic diseases in adult offspring rats fed a high-fat diet after UCS in a gender-specific manner and enhances the susceptibility to metabolic diseases in adult offspring rats fed a high-fat diet.     

Hsp90 inhibitor BILBO21 enhances triptolide-induced apoptosis of human T-cell acute lymphoblastic leukemia cells in vitro mainly by disrupting p53- MDM2 balance

Aim： To investigate the effects of BILBO21, an inhibitor of heat shock protein 90 （Hsp90） alone or in combination with triptolide （TPL） on T-cell acute lymphoblastic leukemia （T-ALL） and the mechanisms of action. Methods： Human T-ALL cells line Molt-4 was examined. The cell viability was measured using M]-I- assay. Apoptotic cells were studied with Hoechst 33258 staining. Cell apoptosis and cell cycle were analyzed using flow cytometry with Annexin V/PI staining and PI staining, respectively. The levels of multiple proteins, including Akt, p65, CDK4/6, p18, Bcl-2 family proteins, MDM2, and p53, were examined with Western blotting. The level of MDM2 mRNA was determined using RT-PCR. Results： Treatment of Molt-4 cells with BILBO21 （50-800 nmol/L） inhibited the cell growth in a dose-dependent manner （the IC~ovalue was 384.6 and 301.8 nmol/L, respectively, at 48 and 72 h）. BILBO21 dose-dependently induced Go/G1 phase arrest, followed by apoptosis of Molt-4 cells. Furthermore, BILBO21 increased the expression of p18, decreased the expression of CDK4/6, and activated the caspase pathway in Molt-4 cells. Moreover, BILBO21 （50-400 nmol/L） dose-dependently decreased the phospho-MDM2 and total MDM2 protein levels, but slightly increased the phospho-p53 and total p53 protein levels, whereas TPL （5-40 nmol/L） dose- dependently enhanced p53 activation without affecting MDM2 levels. Co-treatment with BILBO21 and TPL showed synergic inhibition on Molt-4 cell growth. The co-treatment disrupted p53-MDM2 balance, thus markedly enhanced p53 activation. In addition, the co-treatment increased the expression of Bak and Bim, followed by increased activation of caspase-9. Conclusion： The combination of BILBO21 and TPL may provide a novel strategy for treating T-ALL by overcoming multiple mechanisms of apoptosis resistance.     

美国与欧洲上市后药品安全风险管理进展概述

近10年来,美国食品药品管理局（FDA）、欧洲药品管理局（EMA）及其他相关单位,对上市后药品安全性监测日益重视,在这一领域取得了显著进展,并且制定了一系列新规定。     

Molecular modeling,quantum polarized ligand docking and structure-based 3D-QSAR analysis of the imidazole series as dual AT~ and ETA receptor antagonists

Aim： Both endothelin ETA receptor antagonists and angiotensin AT1 receptor antagonists lower blood pressure in hypertensive patients. A dual AT1 and ETA receptor antagonist may be more efficacious antihypertensive drug. In this study we identified the mode and mechanism of binding of imidazole series of compounds as dual AT1 and ETA receptor antagonists. Methods： Molecular modeling approach combining quantum-polarized ligand docking （QPLD）, MM/GBSA free-energy calculation and 3D-QSAR analysis was used to evaluate 24 compounds as dual AT1 and ETA receptor antagonists and to reveal their binding modes and structural basis of the inhibitory activity. Pharmacophore-based virtual screening and docking studies were performed to identify more potent dual antagonists. Results： 3D-QSAR models of the imidazole compounds were developed from the conformer generated by QPLD, and the resulting models showed a good correlation between the predicted and experimental activity. The visualization of the 3D-QSAR model in the context of the compounds under study revealed the details of the structure-activity relationship： substitution of methoxymethyl and cyclooctanone might increase the activity against AT~ receptor, while substitution of cyclohexone and trimethylpyrrolidinone was important for the activity against ETA receptor; addition of a trimethylpyrrolidinone to compound 9 significantly reduced its activity against AT~ receptor but significantly increased its activity against ETA receptor, which was likely due to the larger size and higher intensities of the H-bond donor and acceptor regions in the active site of ETA receptor. Pharmacophore-based virtual screening followed by subsequent Glide SP, XP, QPLD and MM/GBSA calculation identified 5 potential lead compounds that might act as dual AT1 and ETA receptor antagonists. Conclusion： This study may provide some insights into the development of novel potent dual ETA and AT1 receptor antagonists. As a result, five compounds are found to be the best dual antagonists against AT1R and ETA receptors.     

Sodium hydrosulfide alleviates lung inflammation and cell apoptosis following resuscitated hemorrhagic shock in rats

Aim： To investigate the protective effects of hydrogen sulfide （H2S） against inflammation, oxidative stress and apoptosis in a rat model of resuscitated hemorrhagic shock. Methods： Hemorrhagic shock was induced in adult male SD rats by drawing blood from the femoral artery for 10 min. The mean arterial pressure was maintained at 35-40 mmHg for 1.5 h. After resuscitation the animals were observed for 200 min, and then killed. The lungs were harvested and bronchoalveolar lavage fluid was prepared. The levels of relevant proteins were examined using Western blotting and immunohistochemical analyses. NariS （28 pmol/kg, ip） was injected before the resuscitation. Results： Resuscitated hemorrhagic shock induced lung inflammatory responses and significantly increased the levels of inflammatory cytokines IL-6, TNF-a, and HMGB1 in bronchoalveolar lavage fluid. Furthermore, resuscitated hemorrhagic shock caused marked oxidative stress in lung tissue as shown by significant increases in the production of reactive oxygen species H202 and OH, the translocation of Nrf2, an important regulator of antioxidant expression, into nucleus, and the decrease of thioredoxin I expression. Moreover, resuscitated hemorrhagic shock markedly increased the expression of death receptor Fas and Fas-ligand and the number apoptotic cells in lung tissue, as well as the expression of pro-apoptotic proteins FADD, active-caspase 3, active-caspase 8, Bax, and decreased the expression of Bcl-2. Injection with NariS significantly attenuated these pathophysiological abnormalities induced by the resuscitated hemorrhagic shock. Conclusion： NariS administration protects rat lungs against inflammatory responses induced by resuscitated hemorrhagic shock via suppressing oxidative stress and the Fas/FasL apoptotic signaling pathway.     

树突状细胞的肿瘤个体化治疗

肿瘤免疫治疗这一概念是基于机体的免疫系统在对抗肿瘤时的调节与反应。有效的免疫治疗的最主要的优点在于相对较少的副作用,针对肿瘤细胞的靶向特异性,并且能产生对抗肿瘤特异性抗原的长期记忆[1]。     

人参北芪片的质量控制研究

目的建立人参北芪片的质量标准,控制制剂的质量。方法采用薄层色谱法对制剂中组成药味人参进行鉴别,采用HPLC-蒸发光散射检测法测定制剂中的黄芪甲苷,并进行方法学考察。结果人参的薄层鉴别中,斑点清晰,阴性无干扰。黄芪甲苷在0.9976～14.9640μg与峰面积呈良好的线性关系,平均回收率为98.06%,RSD为1.60%。结论建立的薄层色谱和高效液相色谱定性、定量方法可以用来控制人参北芪片的质量,方法简便可行。     

生长抑素在姑息治疗中的价值

生长抑素是存在于胃黏膜、胰岛、胃肠道神经、垂体后叶和中枢神经系统中的肽激素,在中枢及外周神经系统和包括胰腺、肠道、肾上腺、肾脏和免疫细胞在内的外周组织中广泛分布,它可作用于包括脑、垂体、胰腺、肠道、肾上腺、甲状腺、肺、肾脏和免疫系统等靶器官。     

Oligomannurarate sulfate inhibits CXCL12/SDF-1- mediated proliferation and invasion of human tumor cells in vitro

Aim： JG6 is a novel marine-derived oligosaccharide that has shown to inhibit angiogenesis and tumor metastasis. In this study, we sought to identify the potential target responsible for the anti-cancer activity of JG6. Methods： Human liver cancer cell line Bel-7402 and human cervical cancer cell line HeLa were examined. CXCL12-stimulated cell proliferation and migration were determined using a CCK-8 kit and a transwell assay, respectively. Western blotting was performed to examine the changes in CXCL12/CXCR4 axis. Molecular docking and surface plasmon resonance （SPR） were performed to characterize the possible interaction between JG6 and the CXCL12/CXCR4 axis. Results： Treatment with CXCL12 potently stimulated the proliferation and migration in both Bel-7402 and HeLa cells. Co-treatment of the cells with JG6 （10, 50 and 100 pp=VmL） dose-dependently impeded the CXCL12-stimulated cell proliferation and migration. Furthermore, CXCL12 rapidly induced phosphorylation of AKT, ERK, FAK and Paxillin in Bel-7402 and HeLa cells, whereas pretreatment with JG6 dose-dependently inhibited the CXCL12-induced phosphorylation of these proteins. The SPR assay showed that JG6 bound to CXCL12 with a high affinity. In molecular docking study, JG6 appeared to interact with CXCL12 via multiple polar interactions, including 6 ionic bonds and 7 hydrogen bonds. Conclusion： Inhibition of the CXCL12/CXCR4 axis by JG6 may account for its anticancer activity.     

新生儿缺氧缺血性脑病并发热个案报道

1 病案介绍 患儿,男,新生儿(30 min),因颜面及全身皮肤青紫30 min于2011年1月11日转入儿科.患儿系第一胎,第一产,足月,因"胎儿宫内窘迫,脐绕颈2圈,羊水Ⅱ度污染"于入院前30 min在我院妇产科剖宫产分娩,胎头取出顺利,胎盘完整,无羊水呛入,生后无自主呼吸及心跳,颜面及全身皮肤青紫,肌张力差,插鼻管尚有反应,经心肺复苏,约3 min后心跳恢复,10 min后建立自主呼吸,但呼吸不规则,继续气囊加压给氧辅助呼吸至30 min,自主呼吸明显好转,皮肤青紫有所缓解,拔除气管插管后,患儿未出现呼吸暂停,急转入儿科治疗.其母孕期健康,无特殊服药史,父亲体健,非近亲婚配,无传染病史,无家族性遗传及代谢病史.     

PIK3CA hypomethylation plays a key role in activation of the PI3K/AKT pathway in esophageal cancer in Chinese patients

Aim： To investigate the role of PIK3CA oncogene in tumorigenesis and development of esophageal cancer in Chinese patients at the levels of genetic mutation and epigenetics. Methods： Seventy six esophageal tumor samples and corresponding adjacent normal tissues were collected, and the genomic DNA was extracted. Mutations in the 9th and 20th exons of PIK3CA gene were detected using conventional sequencing. PIK3CA methylation rates in two selected CpG islands （CpG island I and 2） were detected using sub-bisulfate modified sequencing. Pl10~ and pAKT expression levels were detected with Western blotting. Results： In PIK3CA gene of the tumor tissues, G1633C （E545Q） mutation was detected in the 9th exon with a rate of 3.95% （3/76）, whereas mutation was not found in the 20th exon. Nor mutation did occur in PIK3CA gene of the adjacent normal tissues. The methylation rate of the CpG island I had no significant difference between the tumor and adjacent tissues （0.77%±0.009% vs 0.89%±0.008%）, but the methylation rate of the CpG island 2 in the esophageal tumors was significantly lower than that in the adjacent tissues （6.00%±2.80% vs 10.45%±5.51%）. Furthermore, the rate of methylation of the CpG island 2 in TNM stage Ill and IV esophageal cancer （3.84%±2.08%） was significantly lower than in stage I （8.52%±2.55%） and stage II （6.42%±2.36%）. PIK3CA gene hypomethylation in esophageal cancer was significantly correlated with high expression of p110a. Conclusion： PIK3CA gene hypomethylation plays a key role in the tumorigenesis and development of esophageal cancer in Chinese patients, while the mutations of PIK3CA gene have little effect on the development of esophageal cancer.     

Synthesis and bioassay of β-（1,4）-D-mannans as potential agents against Alzheimer＇s disease

Aim： Oligomannurarate 971 derived from a marine plant has shown neuroprotective effects. In this study we synthesized a series of truncated derivatives of the oligosaccharide, and investigated the effect of these derivatives against Aβ peptide toxicity in vitro. Methods： The sulfoxide method was applied to synthesize the derivatives. SH-SY5Y human neuroblastoma cells were treated with Aβ1-40 （2 pmol/L）, and the cell viability was detected using a CCK8 assay. Results： A series of β-（1,4）-D-mannosyl oligosaccharide, ranging from the disaccharide to the hexasaccharide, were synthesized. Addition of 10 pmol/L β-（1,4）-D-mannobiose 6, β-（1,4）-D-mannotriose 9 or β-（1,4）-D-mannotetraose 12 in SH-SY5Y cells significantly attenuated Aβ1-40-induced toxicity. The efficacies were similar to those caused by 10 pmol/L oligomannurarate 971 or alzhemed. Other oligosaccharides including oligomaltoses and oligocelluloses were less active. Conclusion： Synthetic homogeneous short chain β-（1,4）-D-mannans shows neuroprotective effect against Aβ peptide toxicity similar to that of heterogeneous oligomannurarate 971 and alzhemed.     

奥氮平联用舍曲林致皮肤色素沉着及幻觉症状再现1例

1 临床资料 患者,男,32岁.既往体健,否认食物药物过敏史.个人史无特殊,家族史:患者二叔为自杀死亡. 患者于31岁时无诱因慢性起病,表现为认为被议论、被害,凭空听见有人骂自己,于2010年第1次住院,诊断精神分裂症,予奥氮平(最高用至 20 mg/d)系统治疗,3个月后好转出院,坚持服用奥氮平20 mg/d治疗,病情稳定.后因病失去工作,患者开始病情加重,语乱,生活懒散,情绪差,有自杀念头.2011年第2次送入院.     

400例抗菌药物不良反应报告分析

目的探讨该院抗菌药物不良反应(ADR)发生规律及特点,为临床安全用药提供参考。方法对该院2010-2011年收集的400份抗菌药物ADR报告进行回顾性研究,登记ADR名称、级别、结果、药品名称、用药途径、患者年龄、性别、原患疾病。结果 ADR发生集中在40～50岁年龄段,但老年人及儿童发生ADR的比例也较高;其中,静脉滴注297例,占总数74.25%,口服给药96例,占总数24.00%;左氧氟沙星及阿奇霉素ADR发生比例高;ADR临床表现分布广泛,以皮肤及附件损害和消化系统症状为主。结论抗菌药物ADR产生原因较为复杂,广大医务工作者应加强抗菌药物监管,加大对ADR的认识,采取必要干预措施减少或避免ADR的发生。     

DNA aptamers that target human glioblastoma multiforme cells overexpressing epidermal growth factor receptor variant III in vitro

Aim： Aptamers are oligonucleic acid or peptide molecules that bind to a specific target molecule in cells, thus may act as effective vehicles for drug or siRNA delivery. In this study we investigated the DNA aptamers that target human glioblastoma multiforme （GBM） cells overexpressing epidermal growth factor receptor variant III （EGFRvlII）, which was linked to radiation and chemotherapeutic resistance of this most aggressive brain tumor. Methods： A 73-mer ssDNA library containing molecules with 30 nt of random sequence flanked by two primer hybridization sites was chosen as the initial library. Cell systematic evolution of ligands by exponential enrichment （CelI-SELEX） method was used to select the DNA aptamers that target EGFRvlII. The binding affinity of the aptamers was measured using a cell-based biotin-avidin ELISA. Results： After 14 rounds of selection, four DNA aptamers （32, 41, 43, and 47） that specifically bound to the EGFRvlll-overexpressing human glioma U87A cells with K~ values of less than 100 nmol/L were discovered. These aptamers were able to distinguish the U87A cells from the negative control human glioma U87MG cells and HEK293 cells. Aptamer 32 specifically bound to the EGFRvlII protein with an affinity similar to the EGFR antibody （Kd values of aptamer 32 and the EGFR antibody were 0.62±0.04 and 0.32±0.01 nmol/L, respectively）, and this aptamer was localized in the cell nucleus. Conclusion： The DNA aptamers are promising molecular probes for the diagnosis and treatment of GBM.     

利用真实世界数据为决策提供信息

中国的医疗体系正在经历着重大变革。2009年医疗改革措施的启动,标志着全面推进医改的良好开端：向全民提供可负担的医疗。到目前为止,取得的主要成就包括近乎覆盖全民的基本医疗保险、     

Ketanserin improves cardiac performance after myocardial infarction in spontaneously hypertensive rats partially through restoration of baroreflex function

Aim： Baroreflex dysfunction is associated with a higher rate of sudden death after myocardial infarction （MI）. Ketanserin enhances baroreflex function in rats. The present work was designed to examine whether ketanserin improves the post-MI cardiac function and to explore the possible mechanism involved. Methods： Spontaneously hypertensive rats （SHR） were treated with ketanserin （0.3 mg·k-1·d-1）. Two weeks later, blood pressure and baroreflex function were measured, followed by a ligation of the left coronary artery. The expressions of vesicular acetylcholine transporter （VAChT） and a7 nicotinic acetylcholine receptor （a7-nAChR） in ischemic myocardium, angiogenesis, cardiac function, and left ventricular （LV） remodeling were evaluated subsequently. Results： Ketanserin significantly improved baroreflex sensitivity （0.62＋0.21 vs 0.34＋0.12 ms/mmHg, P〈O.01） and vagal tonic activ- ity （heart rate changes in response to atropine, 54.8＋16.2 vs 37.6＋13.4 bpm, P〈O.01） without affecting the blood pressure or basic heart rate in SHR. Treatment of SHR with ketanserin prominently improved cardiac function and alleviated LV remodeling, as reflected by increases in the ejection fraction, fractional shortening, and LV systolic pressure as well as decreases in LV internal diameter and LV relative weight. The capillary density, vascular endothelial growth factor expression, and blood flow in the ischemic myocardium were significantly higher in the ketanserin-treated group. In addition, ketanserin markedly increased the expression of VAChT and （x7-nAChR in ischemic myocardium. Conclusion： Ketanserin improved post-MI cardiac function and angiogenesis in ischemic myocardium. The findings provide a mechanistic basis for restoring baroreflex function using ketanserin in the treatment of Ml.