AgNOR的图像分析在判断子宫内膜癌预后方面的研究

应用图像分析技术对１０３例子宫内膜癌癌细胞中ＡｇＮＯＲ进行了定量研究。结果表明，低分化、肌层浸润＞１／２者的癌细胞核面积、每核ＡｇＮＯＲ颗粒总面积、颗粒数及颗粒平均面积明显大于高分化及无肌层浸润者。Ⅱ、Ⅳ期、复发、死亡患者的癌细胞的核面积及ＡｇＮＯＲ颗粒平均面积较Ⅰ期、无复发、存活者明显增高。ＡｇＮＯＲ的颗粒平均面积多在１．２０μｍ×１．２０μｍ以上，并且ＡｇＮＯＲ颗粒分布多呈聚集型；而预后较好的患者，ＡｇＮＯＲ颗粒平均面积多在亚．１５μｍ×１．１５μｍ左右，颗粒多呈弥散分布。结果提示，细胞内ＡｇＮＯＲ的含量与子宫内膜癌的生物学行为及预后有关，图像分析进行ＡｇＮＯＲ的定量研究较传统的光镜下计数ＡｇＮＯＲ颗粒数目更为客观、准确。     

Dynamic changes of inducible nitric oxide synthase expression in rat’s retina and its role on blood-retinal barrier injury after acute high intraocular pressure

AIM: To clarify the role of inducible nitric oxide synthase (iNOS) in blood-retinal barrier (BRB) injury after acute high intraocular pressure (IOP) in rats. METHODS: Forty-two Sprague-Dawley (SD) rats were randomized into 7 groups [control (Cont), 3, 6, 12, 24, 48, and 72h, n=6]. Except Cont group, other groups’ retina tissue was obtained at corresponding time points after a model of acute high IOP have been established in rats. The expression of iNOS and tight junction protein zonula occludens (ZO)-1 was detected by Western blotting. Evans blue (EB; 3% ) was injected into the great saphenous vein to detect the leakage of EB by spectrophotometer. Nine rats were divided into Cont, 6h, 12h groups, the expression of iNOS was localized by immunofluorescence. In order to verify the role of iNOS in the damage to BRB, thirty-six rats were randomly divided into 4 groups [Cont, Cont+inhibitor (Inh), 6h and 6h+Inh, n=9]. After treatment with the iNOS-specific inhibitor 1400W, the expression of iNOS and ZO-1 and the leakage of BRB were detected again. RESULTS: The immunofluorescence results showed that the expression of iNOS was observed in the Cont group and 6h group, but not in the 12h group. iNOS was mainly expressed in the retinal nerve fiber layer, ganglion cell layer and inner nuclear layer and that it did not colocalize with the retinal ganglion cell marker NeuN but was co-expressed with the vascular endothelial cell marker CD31. Western blotting showed that in the early period (3h, 6h) after acute high IOP, the expression of iNOS was upregulated, then the down-regulation of iNOS were tested in the follow-up timing spots. ZO-1 expression showed a continuous down-regulation after 6h. The quantitative results for EB showed that the amount of EB leakage began to increase at 3h after acute high IOP. At 6h, the leakage of EB was lower, but at 12h, the leakage of EB was highest, after which it gradually recovered but remained higher than that in the Cont group. The expression of iNOS was down-regulated after 1400W treatment. ZO-1 expression was not significantly changed in the Cont+Inh group and the 6h group, and significantly down-regulated in the 6h+Inh group, and the leakage of EB was significantly increased after 1400W treatment. CONCLUSION: These results suggest that the upregulation of iNOS expression in the early stage after acute high IOP may have a protective effect on BRB injury.     

心肌梗死恢复期运动试验和动态心电图检查的意义

37例心肌梗死恢复期患者作运动试验(ET)和动态心电图(AECG)检查,ET 对 ST 段压低的检出率(27%)显著高于 AECG(13.5%);对多支病变两项检查皆有诊断价值;5例两项均有 ST 段压低者3例发生梗死后频发心绞痛,显著多于无 ST 段压低者(P=0.008)。2例发生梗死后急性左心衰者为室壁瘤伴 ET 中 ST 段抬高者,室壁瘤患者两项检查中室性心律失常显著增多。     

Lapatinib increases motility of triple-negative breast cancer cells by decreasing miRNA-7 and inducing Raf-1/MAPK-dependent interleukin-6

Lapatinib, a dual epidermal growth factor receptor (EGFR) and HER2 tyrosine kinase inhibitor (TKI), has been approved for HER2-positive breast cancer patients. Nevertheless, its inhibitory effect on EGFR did not deliver clinical benefits for triple-negative breast cancer (TNBC) patients even EGFR overexpression was frequently found in this disease. Moreover, lapatinib was unexpectedly found to enhance metastasis of TNBC cells, but the underlying mechanisms are not fully understood. In this study, we explored that the level of interleukin-6 (IL-6) was elevated in lapatinib-treated TNBC cells. Treatment with IL-6 antibody abolished the lapatinib-induced migration. Mechanistically, the signaling axis of Raf-1/mitogen-activated protein kinases (MAPKs), c-Jun N-terminal kinases (JNKs), p38 MAPK, and activator protein 1 (AP-1) was activated in response to lapatinib treatment to induce IL-6 expression. Furthermore, our data showed that microRNA-7 directly binds and inhibits Raf-1 3′UTR activity, and that down-regulation of miR-7 by lapatinib contributes to the activation of Raf-1 signaling pathway and the induction of IL-6 expression. Our results not only revealed IL-6 as a key regulator of lapatinib-induced metastasis, but also explored the requirement of miR7/Raf-1/MAPK/AP-1 axis in lapatinib-induced IL-6 expression.     

Safety,Tolerability, and Pharmacokinetics of Intravenous Nemonoxacin in Healthy Chinese Volunteers

Nemonoxacin (TG-873870) is a novel nonfluorinated quinolone with potent broad-spectrum activity against Gram-positive, Gram-negative, and atypical pathogens, including vancomycin-nonsusceptible methicillin-resistant Staphylococcus aureus (MRSA), quinolone-resistant MRSA, quinolone-resistant Streptococcus pneumoniae, penicillin-resistant S. pneumoniae, and erythromycin-resistant S. pneumoniae. This first-in-human study was aimed at assessing the safety, tolerability, and pharmacokinetic properties of intravenous nemonoxacin in healthy Chinese volunteers. The study comprised a randomized, double-blind, placebo-controlled, dose escalating safety and tolerability study in 92 subjects and a randomized, single-dose, open-label, 3-period Latin-square crossover pharmacokinetic study in 12 subjects. The study revealed that nemonoxacin infusion was well tolerated up to the maximum dose of 1,250 mg, and the acceptable infusion rates ranged from 0.42 to 5.56 mg/min. Drug-related adverse events (AEs) were mild, transient, and confined to local irritation at the injection site. The pharmacokinetic study revealed that after the administration of 250, 500, and 750 mg of intravenous nemonoxacin, the maximum plasma drug concentration (C_max) values were 4.826 μg/ml, 7.152 μg/ml, and 11.029 μg/ml, respectively. The corresponding values for the area under the concentration-time curve from 0 to 72 hours (AUC_{0–72 h}) were 17.05 μg · h/ml, 39.30 μg · h/ml, and 61.98 μg · h/ml. The mean elimination half-life (t_1/2) was 11 h, and the mean cumulative drug excretion rate within 72 h ranged from 64.93% to 77.17%. Volunteers treated with 250 to 750 mg nemonoxacin exhibited a linear dose-response relationship between the AUC_{0–72 h} and AUC_0–∞. These findings provide further support for the safety, tolerability, and pharmacokinetic properties of intravenous nemonoxacin. (This study has been registered at ClinicalTrials.gov under registration no. {"type":"clinical-trial","attrs":{"text":"NCT01944774","term_id":"NCT01944774"}}NCT01944774.)     

克林霉素治疗细菌感染60例

目的：评价国产克林霉素磷酸酯的临床疗效和安全性。方法：细菌感染病人６０例（男性３９例，女性２１例；年龄４６±ｓ１７ａ），其中呼吸道感染４９例，尿路感染４例，皮肤软组织感染和急性骨髓炎各３例，败血症１例。用克林霉素磷酸酯１．２～２．４ｇ／ｄ，分２～３次静脉滴注，疗程７～１４ｄ。结果：治疗后临床有效率８５％，痊愈率６２％，细菌清除率９０％，不良反应少而轻微，皮疹和一过性ＡＬＴ升高的发生率分别为５％和２％。结论：国产克林霉素磷酸酯治疗需氧革兰阳性菌感染疗效好且安全。     

甲基莲心碱对氧自由基介导冠状血管阻力变化的影响

研究了甲基莲心碱（Ｎｅｆ）对氧自由基介导离体大鼠冠状血管阻力变化的影响．结果发现电解性氧自由基可引起冠脉灌注压（ＣＡＰＰ）增加；去冠状血管内皮后，电解所致ＣＡＰＰ的增加明显减弱．预先给予Ｌ－精氨酸（１００μｍｏｌ·Ｌ－１）能明显对抗氧自由基所致的ＣＡＰＰ增加，且Ｌ－精氨酸的作用能部分被硝基－Ｌ－精氨酸（２００μｍｏｌ·Ｌ－１）所对抗，提示电解所致ＣＡＰＰ的增加与氧自由基影响冠状血管内皮功能有关．Ｎｅｆ依剂量性地对抗电解所致ＣＡＰＰ的增加，且Ｎｅｆ（１０μｍｏｌ·Ｌ－１）的作用能被硝基－Ｌ－精氨酸（２００μｍｏｌ·Ｌ－１）或吲哚美辛（１μｍｏｌ·Ｌ－１）所对抗，提示Ｎｅｆ的作用可能部分通过影响前列腺素及内皮源性舒张因子类物质的代谢有关．     

The correlation between rat retinal nerve fiber layer thickness around optic disc by using optical coherence tomography and histological measurements

AIM:To explore the correlation between the retinal nerve fiber layer (RNFL) thickness by using optical coherence tomography (OCT) and by histological measurements in normal adult rats and optic nerve transected rats.METHODS:The RNFL thickness of 36 rats was scanned in a circle 3.46mm far from the optic disc by OCT. The two experimental groups were the normal group (n=20 rats) and the optic nerve transected group (n=16 rats). The latter group included 4 groups (n=4/group) surviving for 1 day, 3, 5 and 7 days. Then the RNFL thickness of the same retina area was also measured by NF-200 immunohistochemical staining method. Linear regression was used to analyze the correlation between the data obtained from these two methods.RESULTS: The RNFL thickness of normal right eyes around optic disc by OCT was 72.35±5.71μm and that of the left eyes was 72.65±5.88μm (P=0.074). The RNFL thickness of the corresponding histological section by immunohistochemistry was 37.54±4.05μm (right eyes) and 37.38±4.23μm (left eyes) (P=0.059). There was a good correlation between the RNFL thickness measured by OCT and that measured by histology (R²=0.8131). After optic nerve transection, the trend of the RNFL thickness was thinner with the prolonged survival time. The correlation of the thickness detected by the above two methods was approximately (R²=0.8265). Value of the RNFL thickness in rats around optic disc measured by OCT was obviously higher than that measured by common histological measurement in normal adult rats and optic nerve transected rats.CONCLUSION: The RNFL thickness measured by OCT has a strong correlation with that measured by histological method. Through OCT scanning, we found that the thickness of RNFL gradually becomes thinner in a time-dependent manner.     

2-Phenyl-5-(pyrrolidin-1-yl)-1-(3,4,5-trimethoxybenzyl)-1H-benzimidazole, a benzimidazole derivative, inhibits growth of human prostate cancer cells by affecting tubulin and c-Jun N-terminal kinase

BACKGROUND AND PURPOSE

The c-Jun N-terminal kinase (JNK) and tubulin are, frequently, targets for developing anti-cancer drugs. A major obstacle to successful development is P-glycoprotein (P-gp)-mediated resistance. Here, we have assessed a compound that inhibited growth of cancer cells, for effects on JNK and tubulin and as a substrate for P-gp.

EXPERIMENTAL APPROACH

Several pharmacological and biochemical assays were used to characterize signalling pathways of 2-phenyl-5-(pyrrolidin-1-yl)-1-(3,4,5-trimethoxybenzyl)-1H-benzimidazole (PPTMB), a benzimidazole analogue, in prostate cancer cells.

KEY RESULTS

PPTMB inhibited proliferation of several human prostate cancer cell lines. It displayed similar activity against a P-gp-rich cell line, indicating that PPTMB was not a substrate for P-gp. PPTMB induced G2/M arrest of the cell cycle and subsequent apoptosis, using flow cytometry. Tubulin polymerization assays and Western blot analysis showed that PPTMB directly acted on tubulin and caused disruption of microtubule dynamics, inducing mitotic arrest and sustained high levels of cyclin B1 expression and Cdk1 activation. Subsequently, mitochondria-related apoptotic cascades were induced, including Bcl-2 and Bcl-xL phosphorylation, Mcl-1 down-regulation, truncated Bad formation and activation of caspase-9 and -3. PPTMB stimulated JNK phosphorylation at Thr¹⁸³/Tyr¹⁸⁵. SP600125, a specific JNK inhibitor, significantly inhibited apoptotic signalling, indicating that JNK plays a key role in PPTMB action. PPTMB showed a 10-fold higher potency against prostate cancer cells than normal prostate cells.

CONCLUSIONS AND IMPLICATIONS

PPTMB is an effective anti-cancer agent. It disrupted microtubule dynamics, leading to mitotic arrest of the cell cycle and JNK activation, which in turn stimulated the mitochondria-related apoptotic cascades in prostate cancer cells.