Facile synthesis of photoactivatable adenosine analogs

Adenosine and its derivatives are important building blocks of the biological system. They serve as the universal energy currency, amplify intracellular signals for various signal transduction pathways, and can also be used as the co-substrates for enzymatic transformations. The synthesis and regulation of adenosine and its analogs rely on the adenosine binding proteins (ABPs). Dysregulated ABP activity contributes to numerous diseases such as cancer, metabolic disorders, and neurodegenerative diseases. Presently, there is intense interest in targeting ABPs for therapeutic purposes. A large fraction of the human ABP family remains poorly characterized. The need for innovative chemical probes to investigate ABP function in the native biological matrix is apparent. In this study, an adenosine analog, probe 1, with a photoaffinity group and biotin tag was synthesized using concise synthetic strategies. This probe was able to label and capture individual recombinant ABPs with good target selectivity. Probe 1 was also evaluated for its ability to label spiked ABP in complex cell lysates. This chemical probe, together with the labeling and enrichment assay, is of great value to interrogate the biological functions of ABPs and to elucidate their diversity under different physiological conditions.

Photoactivatable adenosine analog-enabled capture and enrichment of adenosine binding protein (ABP).     

痰热清注射液对产ESBLs肺炎克雷伯菌的体外抑制作用

目的观察产ESBLs肺炎克雷伯菌在体外对中药痰热清注射液以及其与特治星（哌拉西林/他唑巴坦）联合应用的效果,为耐药菌株的治疗提供新的思路。方法采用临床和实验室标准化协会（CLSI）推荐的肉汤稀释法,中药及中西药联合应用对产ESBLs肺炎克雷伯菌进行抑菌试验。结果痰热清注射液对产ESBLs的肺炎克雷伯菌的最小抑菌浓度（MIC）值为750μL/mL;特治星对产ESBLs的肺炎克雷伯菌的MIC值为28.125μg/mL,当痰热清注射液与特治星同时作用于产ESBLs的肺炎克雷伯菌,痰热清注射液浓度为0.001~1.758μL/mL时,可以使特治星的用量比单用时减少1~2倍。结论中西药联合应用具有明显的抑菌效果,可以显著的减少抗菌药物的用量,这对于耐药菌的临床治疗以及减少耐药菌株的产生具有一定的意义。     

外周血癌细胞中多种肿瘤标记物基因检测预测乳腺癌患者转移和预后的意义

目的：筛选一组可检测乳腺癌循环癌细胞的标记物基因,分析乳腺癌患者相对循环癌细胞数（Lc）与临床病理特征的关联。方法：收集乳腺癌患者142例（乳腺癌组）和正常女性60人（正常对照组）外周血标本。利用CGAP提供的SAGE Genie数据库中数字基因表达演示工具,筛选一组可用于检测乳腺癌循环癌细胞的标记物基因（FAM83A、NPY1R和KRT19）。应用定量巢式PCR方法检测研究对象外周血中标记物基因的表达水平,并通过公式计算患者Lc值。结果：在乳腺癌组患者外周血中肿瘤标记物基因FAM83A、NPY1R和KRT19的表达水平均明显高于正常对照组（P<0.001）。142例乳腺癌患者中有113例（79.6%）至少表达1个肿瘤标记物基因。乳腺癌患者Lc值与其临床分期和远处转移有关联（P<0.001）。Kaplam-Meier生存曲线,Lc值小于2的患者生存时间明显长于Lc值大于2者（P=0.005）。结论：筛选了一组检测乳腺癌外周血循环癌细胞的标记物基因,联合该组基因计算的Lc值与患者临床分期和远处转移有关联,并可辅助判断乳腺癌预后。     

SAP-1 is a microvillus-specific protein tyrosine phosphatase that modulates intestinal tumorigenesis

SAP-1 (PTPRH) is a receptor-type protein tyrosine phosphatase (RPTP) with a single catalytic domain in its cytoplasmic region and fibronectin type III-like domains in its extracellular region. The cellular localization and biological functions of this RPTP have remained unknown, however. We now show that mouse SAP-1 mRNA is largely restricted to the gastrointestinal tract and that SAP-1 protein localizes to the microvilli of the brush border in gastrointestinal epithelial cells. The expression of SAP-1 in mouse intestine is minimal during embryonic development but increases markedly after birth. SAP-1-deficient mice manifested no marked changes in morphology of the intestinal epithelium. In contrast, SAP-1 ablation inhibited tumorigenesis in mice with a heterozygous mutation of the adenomatous polyposis coli gene. These results thus suggest that SAP-1 is a microvillus-specific RPTP that regulates intestinal tumorigenesis.     

Significance of Serum Neuron-specific Enolase as a Predictor of Relapse of Small Cell Lung Cancer

We conducted a prospective study to evaluate the significanceof serum neuron-specific enolase (NSE) as a predictor of relapseof small cell lung cancer (SCLC). Patients entered into thestudy were drawn from those who had shown a complete or partialresponse to first-line chemotherapy with a concurrent declinein the NSE level to less than 10 ng/ml. When the serum NSE levelincreased to more than 15 ng/ml, the patient was restaged onthe basis of clinical, radiological, and bronchoscopic examinations.During the period from August 1988 to December 1990, 57 patientswith SCLC were enrolled and followed up until May 1992; Of thesepatients, 45 had clinical relapses, and 14 (31%) of them showeda clear elevation of the serum NSE level prior to the clinicalrecognition of relapse. Although one false-positive case wasnoted, this involved only a transient elevation of the NSE level.In patients who showed increased NSE levels, the relapses occurredin more difficult to detect silent sites such as the adrenalgland, liver, and deep lymph nodes. In addition, the percentageof patients demonstrating high NSE levels who were able to benefitfrom salvage chemotherapy was higher than for those who didnot (P<0.05). Our results indicate that serial NSE measurementsare useful for the early prediction of SCLC relapse and shouldhelp to facilitate early administration of salvage chemotherapyfor affected patients.(P<0.05)     

Relationship between the Pharmacokinetics of Irinotecan and Diarrhea during Combination Chemotherapy with Cisplatin

Two phase I trials of irinotecan (CPT-11) in combination with cisplatin were conducted. In both cases, the dose-limiting toxicities were leukopenia and/or diarrhea. During these trials the pharmacokinetics of CPT-11 and its active metabolite, 7-ethyl-10-hydroxycamptothecin (SN-38), were investigated to evaluate the relationship between pharmacokinetic parameters and diarrhea, since this is an unpredictable and severe toxicity of combination chemotherapy using CPT-11 and cisplatin. Twenty-three previously untreated patients with advanced lung cancer were evaluated in the pharmacokinetic study. Ten patients received CPT-11 at 80 or 90 mg/m² plus cisplatin at 60 mg/m². The other 13 patients received CPT-11 at 80 or 90 mg/m² plus cisplatin at 80 mg/m² with the granulocyte colony-stimulating factor support (2 μg/kg × 16 days). CPT-11 was given as a 90-min intravenous infusion on days 1, 8, and 15. Cisplatin was given on day 1. The pharmacokinetics of CPT-11 and SN-38 were analyzed on day 8 during the first course of treatment. The maximum tolerated dose of CPT-11 was 90 mg/m² in both phase I trials. The severity of diarrhea was best correlated with the peak plasma concentration of SN-38 among the pharmacokinetic parameters tested. In addition, patients with a plasma SN-38 level > 12.4 ng/ml at 1.75 h after the start of CPT-11 infusion had a higher incidence of Eastern Cooperative Oncology Group grade 3–4 diarrhea than those with a lower SN-38 level ( P =0.0003). Stepwise logistic regression analysis identified the SN-38 concentration as a significant contributor to the development of diarrhea ( P =0.0021). We conclude that there is a clear relationship between the SN-38 concentration and diarrhea during chemotherapy with CPT-11 plus cisplatin.     

人眼结膜上皮细胞体外培养治疗翼状胬肉

目的 探讨体外分离培养结膜上皮细胞治疗翼状胬肉的方法.方法 将126例翼状胬肉患者(均为单眼患者)随机分为2组.治疗组58例,术前2周取其穹隆部结膜(1mm×1mm)剪成0．5mm×O．5mm大小后种植于羊膜上体外培养,术中逆行切除胬肉组织,再将培养好的结膜上皮植片移植于结膜创口,缝合固定.对照组68例,单纯切除翼状胬肉,暴露角巩膜缘.两组术后均行妥布霉素眼膏及滴眼液治疗,预防感染.结果 接种在羊膜上的结膜组织,8d左右融合成膜状,细胞为复层上皮细胞.植片3d后开始呈透明或半透明状,1周时可见新生血管长入,植片与周围结膜逐渐融合,2周左右植片与周围结膜完全融合.术后随访1～2年.治疗组复发2例(3．45％),对照组复发23例(33．82％),两组问复发率差异显著,有统计学意义(x2=16．3．P<0．01).所有患者眼球活动自如,未发生睑球粘连、疤痕增生等并发症.结论 体外分离培养结膜上皮细胞治疗翼状胬肉是一种理想的方法.     

Alveolar macrophages from HIV-infected subjects are resistant to Mycobacterium tuberculosis in vitro

HIV-infected individuals frequently develop Mycobacterium tuberculosis (MTB) infection. Alveolar macrophages (AM) are the initial host defense against this organism. We measured MTB growth in AM from normal and HIV-infected subjects after in vitro exposure. Intracellular growth of MTB was reduced in AM from HIV-infected subjects compared with normal macrophages. This was confined to subjects with CD4 counts greater than 200/microl. Growth of avirulent mycobacteria in HIV macrophages was significantly less than virulent MTB. Because avirulent MTB is more sensitive to tumor necrosis factor-alpha (TNF-alpha), we examined the relationship between cytokine secretion and mycobacterial growth. Higher AM spontaneous TNF-alpha secretion was associated with reduced MTB growth in normal AM. This relationship was not seen in HIV-infected subjects, suggesting that other factors contributed to mycobacteria resistance. Mycobacteria-induced TNF-alpha secretion was inversely associated with growth in normal AM but not in HIV-infected subjects. Finally, binding and internalization of MTB was augmented in HIV macrophages compared with normal, demonstrating that reduced intracellular MTB growth was not due to impaired phagocytosis. In conclusion, the increased incidence of MTB infection in HIV-infected subjects does not appear to be due to a defect in macrophage innate immunity.     

Use of automatic threshold tracking function with non-low polarization leads: risk for algorithm malfunction

The AutoCapture (AC) function of new pacemakers (PM) from St Jude Medical (SJM) was originally recommended for use with low polarization (LP) ventricular leads only.However, recent reports have encouraged the use of the AC function with various leads, including those lacking a special LP design. The objective of this study was to analyze the reliability and safety of the AC algorithm application with different types of pacing leads.The study group comprised 30 consecutive patients with AC PMs connected to three different types of non-LP leads. Ten patients with SJM LP leads served as the control group. The study protocol included a complete AC function test using four different pulse widths (PW). The pacing threshold was independently assessed by a manual/semiautomatic check. Erratic behavior of polarization measurements with increasing PWs was demonstrated in 43% (n = 13) of the study group. Invalid polarization measurements resulted in erroneous algorithm recommendation to apply AC function in 17% (n = 5) of the study patients. Subsequent AC function activation lead to incorrect threshold determination due to missed noncapture in three patients. AC function should be applied with caution with non-LP leads. "Off label" use of these leads may cause erroneous polarization signal measurements which, in some cases, may result in incorrect pacing threshold determination, rendering a potential risk to dependent patients.     

Role of pericellular proteolysis by membrane-type 1 matrix metalloproteinase in cancer invasion and angiogenesis

Membrane-type 1 matrix metalloproteinase (MT1-MMP) is an integral membrane proteinase that is frequently expressed in malignant cancer cells and has potent invasion-promoting activity. When expressed on the cell surface, MT1-MMP degrades the extracellular matrix (ECM) barrier adjacent to the cells to maintain the migration route to traverse the tissue. But MT1-MMP is not just an enzyme that degrades ECM. MT1-MMP also introduces limited cleavage into proteins at the cell-ECM interspaces and converts their functions. The target molecules are ECM components, cell adhesion molecules, and latent forms of MMPs. Through these processing events MT1-MMP modulates the migratory and invasive behavior of the cells.