Plasma level monitoring of mitotane (o,p'-DDD) and its metabolite (o,p'-DDE) during long-term treatment of cushing's disease with low doses

Summary. Mitotane (o,p'-DDD) can be used for the treatment of various adrenocortical diseases such as Cushing's syndrome, but the usual doses of 6–8 g per day are often associated with severe adverse effects.This paper reports the results of much lower doses of o,p'-DDD (0.5–2 g per day) in two patients with Cushing's disease over periods of 8 and 5 years, respectively, under concomitant monitoring of the plasma levels of the parent drug and its major metabolite, o,p'-DDE.It became apparent that o,p'-DDD and o,p'-DDE have a strong tendency to accumulate in the body due to their high lipophilicity. As a consequence, changes in dose regimens had long lag times before they were reflected in plasma levels and once an increase or decrease had started one had to be careful not to cause overshoot.Steady state plasma levels of o,p'-DDD between 5–10 g/ml appeared sufficient to induce and to maintain remission of the disease, which was accompanied with normal cortisol levels in plasma and urine. DDD-levels below 5 g/ml for several weeks may lead to relapses, whereas DDD-levels over 10 g/ml gave rise to side effects. On the other hand, o,p'-DDE seemed inactive at levels up to 4 g/ml in plasma.     

原发性干燥综合征患者B细胞活化因子受体的表达

目的 检测原发性干燥综合征(pSS)患者血清中B细胞活化因子受体(BAFF-R)的表达,探讨其在pSS发病中的作用.方法 pSS患者120例,采用酶联免疫吸附法(ELISA)检测血清BAFF-R水平,对照组62例为健康体检者,性别、年龄构成与患者组匹配.分析血清BAFF-R表达与免疫指标、炎性指标及自身抗体的相关性.结果 血清BAFF-R平均水平pSS组为(692.7±536.9)ng/L,对照组为(279.7±186.9)ng/L,pSS组明显高于对照组(P＜0.01).血清BAFF-R水平与免疫球蛋白IgG呈正相关(r=0.429,P＜0.01),与免疫球蛋白IgA、IgM、补体C3、C4、类风湿因子、红细胞沉降率、C反应蛋白无明显相关性.抗核抗体、抗SSA抗体、抗SSB抗体阳性者血清BAFF-R水平高于阴性者.随着抗核抗体滴度的增加,血清BAFF-R水平呈平行增高趋势.结论 pSS患者中BAFF-R表达增高,并与免疫球蛋白定量、自身抗体表达呈相关性.BAFF-R的异常表达参与了pSS的发病.     

来氟米特联合间歇小剂量环磷酰胺治疗原发性干燥综合征合并间质性肺疾病八例

目的探讨来氟米特(LEF)联合间歇小剂量环磷酰胺(CTX)治疗原发性干燥综合征(pSS)合并间质性肺疾病(ILD)的有效性及安全性。方法给予8例pSS合并ILD患者LEF(10 mg/d)联合间歇小剂量CTX(200~400 mg/3周)治疗,随访1~2.5年,观察8例患者治疗前后的临床转归及实验室检查改变。结果8例患者口眼干燥及干咳、劳力性气促症状明显减轻,肺功能有所恢复,肺总量(TLC)、一氧化碳弥散度(DLco)均有不同程度的增加,肺高分辨率CT病变面积减少,但肺底吸气末期Velcro音变化不明显。无明显不良反应发生。结论LEF联合间歇小剂量CTX治疗pSS合并ILD是有效、安全、可行的。     

毛果芸香碱治疗原发性干燥综合征44例疗效观察

目的观察毛果芸香碱治疗原发性干燥综合征所致口干的疗效和不良反应。方法44名原发性干燥综合征患者参加本随机双盲对照试验。患者随机接受4mg毛果芸香碱或16mg必嗽平片剂。每天4次,共28d。通过对主观口干症状评价和唾液量分泌量测定方式进行全面评估。结果与对照组相比,毛果芸香碱组口干不适症状获得显著改善、唾液分泌量明显增加。结论表明毛果芸香碱治疗干燥综合征口干有效、安全,而且耐受性良好。     

Guadecitabine (SGI-110): an investigational drug for the treatment of myelodysplastic syndrome and acute myeloid leukemia

ABSTRACT

Introduction: The incidence of acute myeloid leukemia (AML) and myelodysplastic syndromes (MDS) is increasing with the aging population. Prognosis and overall survival (OS) remain poor in elderly patients and in those not eligible for intensive treatment. Hypomethylating agents (HMAs) have played an important role in this group of patients but their efficacy is limited.

Areas covered: This article reviews the mechanism of action, pharmacology, safety profile and clinical efficacy of subcutaneous guadecitabine, a second-generation DNA methylation inhibitor in development for the treatment of AML and MDS.

Expert opinion: Although guadecitabine did not yield improved complete remission (CR) rates and OS compared to the control arm in patients with treatment-naïve AML who were ineligible for intensive chemotherapy, subgroup analysis in patients who received ≥4 cycles of therapy demonstrated superior outcomes in favor of guadecitabine. Given its stability, ease of administration, safety profile and prolonged exposure time, guadecitabine would be the more appropriate HMA, replacing azacitidine and decitabine, to be used combination treatment regimens in patients with myeloid malignancies.     

NP-313, 2-acetylamino-3-chloro-1,4-naphthoquinone, a novel antithrombotic agent with dual inhibition of thromboxane A(2) synthesis and calcium entry

BACKGROUND AND PURPOSE

1,4-Naphthoquinones exhibit antiplatelet activity both in vivo and in vitro. In the present study, we investigated the antiplatelet effect of a novel naphthoquinone derivative NP-313, 2-acetylamino-3-chloro-1,4-naphthoquinone and its mechanism of action.

EXPERIMENTAL APPROACH

We measured platelet aggregation, Ca²⁺ mobilization, thromboxane B2 formation and P-selectin expression and examined several enzymatic activities. Furthermore, we used the irradiated mesenteric venules in fluorescein sodium–treated mice to monitor the antithrombotic effect of NP-313 in vivo.

KEY RESULTS

NP-313 concentration-dependently inhibited human platelet aggregation induced by collagen, arachidonic acid, thapsigargin, thrombin and A23187. NP-313 also inhibited P-selectin expression, thromboxane B₂ formation and [Ca²⁺]_i elevation in platelets stimulated by thrombin and collagen. NP-313 at 10 µM inhibited cyclooxygenase, thromboxane A₂ synthase, and protein kinase Cα, whereas it did not affect phospholipase A₂ or phospholipase C activity. In the presence of indomethacin and an adenosine 5-diphosphate scavenger, NP-313 concentration-dependently inhibited thrombin- and A23187-induced [Ca²⁺]_i increase through its inhibitory effects on Ca²⁺ influx, rather than blocking Ca²⁺ release from intracellular stores. NP-313 also inhibited thapsigargin-mediated Ca²⁺ influx through store-operated calcium channel but had no effect on Ca²⁺ influx through store-independent calcium channel evoked by the diacylglycerol analogue 1-oleoyl-2-acetyl-sn-glycerol. Nevertheless, it had little effect on cyclic AMP and cyclic GMP levels. Also, intravenously administered NP-313 dose-dependently inhibited the thrombus occlusion of the irradiated mesenteric vessels of fluorescein-pretreated mice.

CONCLUSIONS AND IMPLICATIONS

Taken together, these results indicate that NP-313 exerts its antithrombotic activity through dual inhibition of thromboxane A₂ synthesis and Ca²⁺ influx through SOCC.     

Renzapride: a new drug for the treatment of constipation in the irritable bowel syndrome

Renzapride is a novel drug currently under clinical evaluation for the treatment of irritable bowel syndrome (IBS). Renzapride is a mixed 5-hydroxytryptamine type 4 (5-HT₄) agonist and 5-HT₃ receptor antagonist that has a stimulatory effect on gastrointestinal motility and transit, as established by in vivo and in vitro studies. Its therapeutic efficacy, tolerability and safety have been evaluated in diabetic gastroparesis in a single study, as well as in IBS in a few other studies. Phase II studies indicated potential beneficial effects on symptoms and bowel habits in patients with constipation-predominant IBS and mixed-type IBS. The outcome of Phase III studies is currently under evaluation.     

Design, synthesis and in vitro PDE4 inhibition activity of certain quinazolinone derivatives for treatment of asthma

In this study, a novel series of quinazolinone derivatives analogue to nitraquazone structure were synthesized. The compounds tested for their inhibitory activity against phosphodiesterase 4B revealed that compound 6d shows promising inhibitory activity comparable to that of Rolipram, whereas compounds 6a and 6c exhibited moderate inhibitory activity.     

Double blockade of angiotensin II (AT(1) )-receptors and ACE does not improve weight gain and glucose homeostasis better than single-drug treatments in obese rats

BACKGROUND AND PURPOSE

Combination therapies are becoming increasingly important for the treatment of high blood pressure. Little is known about whether double blockade of angiotensin II (AT₁) receptors and angiotensin-converting enzyme (ACE) exert synergistic metabolic effects.

EXPERIMENTAL APPROACH

Spontaneously hypertensive rats were allowed to choose between palatable chocolate bars and standard chow and were simultaneously treated with the AT₁ blocker telmisartan (8 mg·kg_bw⁻¹·day⁻¹), the ACE inhibitor ramipril (4 mg·kg_bw⁻¹·day⁻¹) or a combination of the two (8 + 4 mg·kg_bw⁻¹·day⁻¹) for 12 weeks.

KEY RESULTS

Although food-dependent energy intake was increased by telmisartan and telmisartan + ramipril compared with ramipril or controls, body weight gain, abundance of fat and plasma leptin levels were decreased. Increased insulin levels in response to an oral glucose tolerance test were comparably attenuated by telmisartan and telmisartan + ramipril, but not by ramipril. During an insulin tolerance test, glucose utilization was equally as effectively improved by telmisartan and telmisartan + ramipril. In response to a stress test, ACTH, corticosterone and glucose increased in controls. These stress reactions were attenuated by telmisartan and telmisartan + ramipril.

CONCLUSIONS AND IMPLICATIONS

The combination of telmisartan + ramipril was no more efficacious in regulating body weight and glucose homeostasis than telmisartan alone. However, telmisartan was more effective than ramipril in improving metabolic parameters and in reducing body weight. The association between the decrease in stress responses and the diminished glucose levels after stress supports our hypothesis that the ability of telmisartan, as an AT₁ receptor blocker, to alleviate stress reactions may contribute to its hypoglycaemic actions.     

Suppression of apoptosis and induction of DNA synthesis in vitro by the phthalate plasticizers monoethylhexylphthalate (MEHP) and diisononylphthalate (DINP): a comparison of rat and human hepatocytes in vitro

Diethylhexylphthalate (DEHP) and diisononylphthalate (DINP) are plasticizers with many important commercial, industrial and medical applications. However, both DEHP and DINP are rodent peroxisome proliferators (PPs), a class of compounds that cause rodent liver tumours associated with peroxisome proliferation, induction of hepatic DNA synthesis and the suppression of apoptosis. Despite these effects in the rodent, humans appear to be nonresponsive to the adverse effects of PPs. Previously, we have shown that the fibrate hypolipidaemic peroxisome proliferator, nafenopin, induced DNA synthesis and suppressed apoptosis in rat but not in human hepatocytes. In this work, we have examined species differences in the response of rat and human hepatocytes to DEHP and DINP in vitro. In rat hepatocytes in vitro, both DINP and MEHP (a principle metabolite of DEHP and the proximal peroxisome proliferator) caused a concentration-dependent induction of DNA synthesis and suppression of both spontaneous and transforming growth factor β1 (TGFβ1)-induced apoptosis. Similarly, both MEHP and DINP caused a concentration-dependent induction of peroxisomal β-oxidation although the response to DINP was less robust. In contrast to the pleiotropic response noted in rat hepatocytes, neither DINP nor MEHP caused an induction of β-oxidation, stimulation of DNA synthesis and suppression of apoptosis in human hepatocytes cultured from three separate donors. These data provide evidence for species differences in the hepatic response to the phthalates DEHP and DINP, confirming that human hepatocytes appear to be refractory to the hepatocarcinogenic effects of PPs first noted in rodents. Received: 16 August 1999 / Accepted: 21 September 1999     

Three-dimensionally engineered biomimetic tissue models for in vitro drug evaluation: delivery,efficacy and toxicity

Introduction: Three-dimensionally (3D) engineered biomimetic tissue models are sought after due to their high fidelity in mimicking various native tissues of the human body, this quality of which gives them an important role at the forefront of drug discovery and development. A multitude of studies have consistently indicated that gene expression profiles, cellular phenotypes, differentiation capabilities and functionalities are all affected by tissue architecture. Thus, the drug evaluation process will stand to gain immense benefits from the fairly accurate predictions of cellular responses displayed by 3D-engineered tissue models when exposed to the drugs of interest in vitro. Stemming from this fact, many studies have set out to capitalize on developing tissue models that are tailored to specific aspects of drug evaluation including the tests of novel drug delivery systems, drug efficacy and toxicity.

Areas covered: The areas covered include fabrication methods and usage of 3D in vitro tumor models in cancer research, focusing on the evaluation of delivery and efficacy of various anticancer drugs or other therapeutic agents. Also covered are the use of 3D in vitro inflammatory tissue models in anti-inflammation research, centering on osteoarthritis (OA) and rheumatoid arthritis (RA) and the use of 3D in vitro tissue models designed for drug toxicity evaluation specifically with liver-mimetic tissues.

Expert opinion: Currently available 3D tissue models in various fields of research have already displayed their capabilities in predicting cellular responses to various therapeutic agents and delivery methods with better accuracy than their 2D counterparts, albeit being in need of much refinement before they can be successfully applied for reliable drug evaluation. Given further development and improvement, it is highly probable that the 3D-engineered tissue models may perform as living platforms for dynamic drug evaluation in vitro.     

Effect of Linum usitatissimum L. (linseed) oil on mild and moderate carpal tunnel syndrome: a randomized,double-blind,placebo-controlled clinical trial

Background

Carpal tunnel syndrome is known as the most common entrapment neuropathy. Conservative treatments cannot reduce the symptomatic severity satisfactorily; therefore, effectiveness of Linum usitatissimum L. (linseed) oil on carpal tunnel syndrome, as a complementary treatment, was evaluated in the current study. Linseed oil is a well-known preparation in Iranian traditional medicine and its analgesic, anti-inflammatory and anti-oxidative effects have been shown in previous studies.

Methods

A randomized, double-blind, placebo-controlled clinical trial was conducted. One hundred patients (155 hands) with idiopathic mild to moderate carpal tunnel syndrome aged between 18 and 65 years old were randomized in two parallel groups. These two groups were treated during 4 weeks with topical placebo and linseed oil. In addition, a night wrist splint was prescribed for both groups. Symptomatic severity and functional status were measured using Boston Carpal Tunnel Questionnaire. In addition, median sensory nerve conduction velocity, motor distal latency, sensory distal latency and compound latency as electrodiagnostic parameters were measured at baseline and after the intervention period.

Results

After the intervention, significant improvement was observed regarding Boston Carpal Tunnel Questionnaire symptomatic severity and functional status mean differences (p <0.001) in the linseed oil group compared with those in the placebo group. Also, regarding the mean differences of both groups, significant improvement of nerve conduction velocity of the median nerve was seen in the linseed oil group by a value of 2.38 m/sec (p < 0.05). However, motor distal latency and sensory distal latency of the median nerve showed no between-group significant changes (p = 0.14 for both items). Finally, compound latency was improved slightly in the case group, comparing mean differences between the groups (p <0.05). No significant adverse events were reported from using linseed oil.

Conclusions

It seems that linseed oil could be effective in the management of mild and moderate carpal tunnel syndrome, especially in improving the severity of symptoms and functional status. In addition, its effect on electerodiagnostic parameters, especially on the nerve conduction velocity, can be considered as a valuable point.     

Jack of all trades: Pleiotropy and the application of chemically modified tetracycline-3 in sepsis and the acute respiratory distress syndrome (ARDS)

Sepsis is a disease process that has humbled the medical profession for centuries with its resistance to therapy, relentless mortality, and pathophysiologic complexity. Despite 30 years of aggressive, concerted, well-resourced efforts the biomedical community has been unable to reduce the mortality of sepsis from 30%, nor the mortality of septic shock from greater than 50%. In the last decade only one new drug for sepsis has been brought to the market, drotrecogin alfa-activated (Xigris™), and the success of this drug has been limited by patient safety issues. Clearly a new agent is desperately needed. The advent of recombinant human immune modulators held promise but the outcomes of clinical trials using biologics that target single immune mediators have been disappointing. The complex pathophysiology of the systemic inflammatory response syndrome (SIRS) is self-amplifying and redundant at multiple levels. In this review we argue that perhaps pharmacologic therapy for sepsis will only be successful if it addresses this pathophysiologic complexity; the drug would have to be pleiotropic, working on many components of the inflammatory cascade at once. In this context, therapy that targets any single inflammatory mediator will not adequately address the complexity of SIRS. We propose that chemically modified tetracycline-3, CMT-3 (or COL-3), a non-antimicrobial modified tetracycline with pleiotropic anti-inflammatory properties, is an excellent agent for the management of sepsis and its associated complication of the acute respiratory distress syndrome (ARDS). The purpose of this review is threefold: (1) to examine the shortcomings of current approaches to treatment of sepsis and ARDS in light of their pathophysiology, (2) to explore the application of COL-3 in ARDS and sepsis, and finally (3) to elucidate the mechanisms of COL-3 that may have potential therapeutic benefit in ARDS and sepsis.     

重症急性呼吸综合征22例临床特点及综合防治方法

目的 探讨重症急性呼吸综合征(SARS)的临床特点和综合防治方法。方法 对2003年2月至5月我院收治的22例SARS病人的临床表现、X线特征、实验室资料、综合防治方法等进行分析。结果 发病2周内有密切接触SARS病人的16例；宰杀过动物的2例；潜伏期3～12d。所有病人均以发热为首发症状，其中有高热(39℃)以上者占91％。伴呼吸困难的68％，咳嗽86，4％，乏力72．7％．全身酸痛27％，头痛50％，头晕22．7％，胸痛31．8％，呼吸道卡他症状13．6％．心悸36．4％，腹泻18．2％，血丝痰9％，畏寒18．2％，咽痛9％。77％的病人肺部无明显体征。X线胸片及肺部CT：单侧病灶36．4％，双侧63．6％，双肺弥漫性渗出22．7％。ARDS2例；死亡1例。死亡率4．5％。规律应用皮质类固醇治疗的占72．7％。应用无创正压通气(NIPPV)的5例；应用有创机械通气的2例。通过规范的防护措施，一线医务人员感染率为零；医务人员家属感染率为零；院内交叉感染率为零。结论 综合治疗方案中，合理应用糖皮质激素是重要措施。重症病人应及早使用机械通气。规范的防护措施可预防感染。     

Easy and convenient millimole-scale synthesis of new,potential biomarkers for gamma-hydroxybutyric acid (GHB) intake: Feasible for analytical laboratories

New biomarkers indicating the abuse of drugs and alcohol are still of major interest for clinical and forensic sciences. The endogenous neurotransmitter and approved drug, gamma-hydroxybutyric acid (GHB), is often illegally used for drug-facilitated crimes by spiking GHB into alcoholic beverages. Analytical detection windows of only 6 h in blood and 12 h in urine are often too short to provide reliable proof of GHB ingestion. Therefore, new biomarkers are needed to prove exogenous GHB administration. Previously, amino acid GHB conjugates were discovered in an untargeted metabolomics screening and fatty acid esters with GHB were recently discussed as promising biomarkers to enlarge the analytical detection time windows. However, the development of analytical methods is still slowed down since reference compounds for targeted screenings are still missing. In this paper, we describe simple procedures for the rapid synthesis and purification of amino acid GHB conjugates as well as fatty acid esters, which can be adopted in analytical and clinical/forensic laboratories. Structural characterization data, together with IR, ¹H-nuclear magnetic resonance (NMR), ¹³C-NMR, high-resolution mass spectra (MS), and MS/MS spectra in positive and negative ionization mode are reported for all obtained GHB conjugates and GHB conjugate precursors.     

纳洛酮治疗急性新生儿呼吸窘迫综合征的临床疗效及其心肌保护作用

目的 探讨纳洛酮对急性新生儿呼吸窘迫综合征（NRDS）的临床疗效及心肌的保护作用。方法 入选急性NRDS100例,随机分为对照组（50例）和研究组（50例）;对照组给予常规治疗,研究组在对照组治疗的基础上加用纳洛酮治疗。比较两组治疗前后血清肌酸激酶（CK）、肌酸激酶同工酶（CK-MB）和肌钙蛋白I （cTnI）水平。比较两组住院天数、机械通气天数、用氧天数、呼吸机参数、氧合功能指标[动脉氧分压（PaO₂）、氧合指数（OI）及动脉血/肺泡气氧分压比（a/APO₂）]及治愈率。结果 治疗后研究组CK、CK-MB、cTnI水平均显著低于对照组,差异有统计学意义（P<0.01）。研究组氧合功能各指标均优于对照组,差异有统计学意义（P<0.05）。研究组呼吸机参数除PEEP外均优于对照组,差异有统计学意义（P<0.05）。研究组的用氧天数、机械通气天数、住院天数均低于对照组,差异有统计学意义（P<0.05）。两组治愈率相比差异无统计学意义。结论 纳洛酮有助于提高NRDS整体疗效,对NRDS心肌保护作用明显。     

Intracellular mechanisms mediating the inhibition of apoB-containing lipoprotein synthesis and secretion in HepG2 cells by avasimibe (CI-1011), a novel acyl-coenzyme A: cholesterol acyltransferase (ACAT) inhibitor

We have studied the cellular and molecular mechanisms involved in the suppression of apoB secretion from HepG2 cells following incubation with avasimibe (CI-1011), a novel inhibitor of acyl-coenzyme A: cholesterol acyltransferase (ACAT). Cellular lipid analysis revealed that avasimibe significantly decreased the synthesis of cholesterol and cholesteryl ester, and, at higher doses, of triglyceride. Time-course trypsin protection assays revealed that avasimibe induced the accumulation of translocationally arrested apoB intracellularly. Pulse-chase studies showed that the treatment with avasimibe induced a >75% decrease in apoB secretion relative to control, but initially enhanced the protein stability and cellular accumulation of apoB. Subcellular fractionation of microsomes further confirmed the accumulation of secretion-incompetent apoB-lipoproteins in the endoplasmic reticulum (ER) and Golgi compartments of avasimibe-treated HepG2 cells. Although incubation of drug-treated cells with carbobenzoxyl-leucinyl-leucinyl-leucinal (MG132), a potent proteasome inhibitor, increased cellular apoB (70%), it failed to increase apoB secretion. Drug treatment induced an accumulation of secretion-incompetent apoB-containing lipoprotein particles, the majority of which demonstrated a density in a range similar to that of high-density lipoprotein. However, studies in permeabilized cells demonstrated that, at longer chase times, intracellularly accumulated apoB was eventually degraded, indicating that the inhibition of degradation may be transient. Oleate treatment of avasimibe-treated cells partially restored apoB secretion but not to the levels seen in control cells. In summary, we hypothesize that avasimibe acutely blocks the secretion of apoB and its associated lipoproteins from HepG2 cells, transiently enhancing its membrane association and cellular accumulation with eventual intracellular degradation of accumulated apoB.     

原发性干燥综合征并发非霍奇金淋巴瘤患者唇腺组织BCL-6基因重排的检测及意义

目的 探讨原发性干燥综合征（pSS）并发非霍奇金淋巴瘤（NHL）患者唇腺组织BCL-6基因重排的临床病理意义.方法 将研究对象分为pSS并发NHL组（16例）,pSS未并发NHL组（20例）,NHL未并发pSS组（阳性对照组,20例）,健康人群组（阴性对照组,20例）,采用荧光原位杂交技术检测4组研究对象唇腺组织BCL-6基因的重排情况.结果 pSS并发NHL组的BCL-6基因重排发生率为56.2％（9/16）,pSS未并发NHL组的BCL-6基因重排发生率为20.0％（4/20）,NHL未并发pSS组的BCL-6基因重排发生率为50.0％（10/20）,正常人群组的BCL-6基因重排发生率为5.0％（1/20）.4组BCL-6基因重排发生率比较差异有统计学意义（χ2=-84.225,P=-0.000）.pSS并发NHL组与NHL未并发pSS组BCL-6基因重排发生率比较差异无统计学意义（χ2=2.251,P=0.139）.pSS并发NHL组BCL-6基因重排发生率显著高于pSS未并发NHL组（χ2=78.658,P=0.000）及健康人群组（χ2=88.391,P=0.000）.结论 BCL-6基因重排与pSS并发NHL相关,BCL-6基因重排可作为预测pSS并发NHL的生物学参考指标.