Evaluation of the vascular architecture of hepatocellular carcinoma by micro flow imaging: pathologic correlation.

OBJECTIVE: The purpose of this study was to evaluate micro flow imaging (MFI) in depicting the vascular architecture of hepatocellular carcinoma (HCC) and the correlation between pathologic differentiation and the intratumoral vascular architecture pattern. METHODS: Micro flow imaging and contrast harmonic imaging (CHI) were performed in 37 patients with HCC. A sulfur hexafluoride-filled microbubble contrast agent was used. The enhancement level and intratumoral vessels were evaluated on CHI. The vascular architecture of each tumor was evaluated on MFI. Pathologic differentiation of the tumors was classified according to the Edmondson grading system. RESULTS: All 37 HCCs showed hyperenhancement in the arterial phase and hypoenhancement in the portal and late phases on CHI. Intratumoral vessels in the arterial phase were visualized in 20 (54.1%) HCCs. On MFI, the vascular architecture in all lesions was clearly delineated and categorized into 3 patterns: cotton, shrubbery, and deadwood, identified in 12 (32.4%), 22 (59.5%), and 3 (8.1%) of the tumors evaluated, respectively. A chi(2) test showed that pathologic differentiation significantly correlated to the vascular pattern (P = .006). Three (75%) of 4 Edmondson grade I HCCs showed the cotton pattern; 18 (75.0%) of 24 Edmondson grade II HCCs showed the shrubbery pattern; and the deadwood pattern was shown only in Edmondson grade III and IV HCCs. CONCLUSIONS: The MFI technique is more effective in depicting the intratumoral vascular architecture. The vascular architecture pattern correlates with pathologic differentiation of HCC.     

Nesfatin-1 Decreases Excitability of Dopaminergic Neurons in the Substantia Nigra

Nesfatin-1, a newly discovered satiety molecule which reduces feeding behavior, has been recognized as a unique regulatory neuropeptide with its multiple roles, both central and peripheral. However, whether it had neuronal modulation effect on dopaminergic neurons is largely unknown. In the present study, using whole-cell patch clamp under current-clamp mode, we investigate the effects of nesfatin-1 on the electrical activity of rat nigral dopaminergic neurons. Nesfatin-1 could produce a resting membrane potential hyperpolarization on the majority of dopaminergic neurons tested. The spike frequency decreased by 23.13?±?5.93 and 43.20?±?5.56 % in 5-nM and 10-nM nesfatin-1 groups, respectively. These effects persisted in the presence of ionotropic glutamate and GABA receptor antagonists. Our study suggests that nesfatin-1 postsynaptically inhibits the electrical activity of nigral dopaminergic neurons.     

纤维支气管镜刷片在肺癌诊断中的应用

在纤维支气管镜直视下将可疑部位或病变部位直接刷片,做细胞学检查。结果:(1)纤维支气管镜刷片阳性率为71.2%,中央型肺癌阳性检出率为96.5%,明显高于上组(P<0.01)。(2)纤维支气管镜刷片分型符合率78.1%,其中小细胞癌95.5%,显著大于鳞癌65.3%和腺癌62.2%(P<0.05)。纤维支气管镜刷片对小细胞癌与中央型肺癌具有很高的应用价值。     

胆管癌组织中信号转导及转录激活因子3信号通路相关基因的表达与意义

目的 探讨胆管癌组织中信号转导及转录激活因子3（STAT3）信号通路相关基因Survivin及环氧化酶2（COX-2）的表达及其与胆管癌患者临床病理特征和预后的关系.方法 收集2007年9月至2012年7月安徽医科大学附属省立医院收治的43例胆管癌患者的癌组织标本,收集同期行胆肠内引流术和胆管修复的12例肝内外胆管结石和胆管损伤患者的正常胆管组织标本作为对照.应用免疫组织化学染色检测胆管癌及正常胆管组织中STAT3、磷酸化STAT3（p-STAT3）、Survivin及COX-2蛋白的表达,并分析其与胆管癌患者临床病理特征和预后的关系.采用门诊或电话方式对胆管癌患者进行随访,随访时间截至2014年3月.计数资料采用x2检验,相关性分析采用Spearman检验.采用Kaplan-Meier法绘制生存曲线,生存分析采用Log-rank检验.结果 STAT3、p-STAT3、Survivin、COX-2蛋白在胆管癌组织中阳性表达率分别为69.8％ （30/43）、65.1％（28/43）、72.1％ （31/43）、79.1％（34/43）,在正常胆管组织中阳性表达率分别为41.7％（5/12）、8.3％（1/12）、16.7％（2/12）、41.7％（5/12）.p-STAT3、Survivin、COX-2蛋白在胆管癌组织中的阳性表达率显著高于正常胆管组织,两组比较,差异有统计学意义（x2=12.136,9.811,4.679,P＜0.05）.p-STAT3、Survivin、COX-2蛋白在胆管癌组织中的高表达与肿瘤局部浸润转移有关（x2=14.700,5.959,4.075,P＜0.05）;p-STAT3蛋白阳性表达与肿瘤神经侵犯有关（x2=10.384,P＜0.05）;Survivin、COX-2蛋白表达与胆管癌患者肿瘤神经侵犯无关（x2=2.718,3.024,P＞0.05）;p-STAT3、Survivin、COX-2蛋白表达与胆管癌患者性别、年龄、肿瘤位置、分化程度、肿瘤直径无关（x2=0.148,0.720,1.835,1.040,0.236;0.001,0.009,0.029,1.863,0.197;0.433,0.686,0.002,2.974,0.029,P＞0.05）.Survivin及COX-2蛋白的表达与p-STAT3呈正相关（r=0.524,0.583,P＜0.05）.43例胆管癌患者获得随访,随访时间为6个     

A novel reference coordinate system to locate the inferomedial point of the transverse-sigmoid sinus junction

Background

A coordinate system was previously developed to identify landmarks on the skull surface to help locate the transverse-sigmoid sinus junction in order to reduce surgical morbidity in retrosigmoid craniotomy; however, in practice we found that this system has important flaws.

Objective

To develop and evaluate a novel reference coordinate system to precisely locate the inferomedial point of the transverse-sigmoid sinus junction (IMTS) and evaluate the effect of gender and skull side (left or right).

Methods

Forty-two adult skulls (84 sides) were obtained for analyses. The X-axis was defined by point A (where the upper edge of the zygomatic arch joins with the frontal process of the zygomatic bone) and point B (where the upper edge of the zygomatic arch blends posterosuperiorly into the supramastoid crest). The Y-axis was defined by the line perpendicular to the X-axis and extending across the tip of the mastoid. The x and y coordinates of IMTS (IMTS-x and IMTS-y) were measured in this coordinate system.

Results

There were 20 male skulls and 22 female skulls. The mean IMTS-x measurements were significantly higher on the right side compared with the left side in both males and females. For the left skull side, the mean IMTS-y measurements were significantly lower in females compared with males.

Conclusion

This novel reference coordinate system may be a reliable and practical method for identifying the IMTS during retrosigmoid craniotomy. There are significant differences in location of the axes with regard to gender and skull side.     

Targeted Disruption of G0/G1 Switch Gene 2 Enhances Adipose Lipolysis,Alters Hepatic Energy Balance,and Alleviates High-Fat Diet–Induced Liver Steatosis

Recent biochemical and cell-based studies identified G₀/G₁ switch gene 2 (G0S2) as an inhibitor of adipose triglyceride lipase (ATGL), a key mediator of intracellular triacylglycerol (TG) mobilization. Here, we show that upon fasting, G0S2 protein expression exhibits an increase in liver and a decrease in adipose tissue. Global knockout of G0S2 in mice enhanced adipose lipolysis and attenuated gain of body weight and adiposity. More strikingly, G0S2 knockout mice displayed a drastic decrease in hepatic TG content and were resistant to high-fat diet (HFD)-induced liver steatosis, both of which were reproduced by liver-specific G0S2 knockdown. Mice with hepatic G0S2 knockdown also showed increased ketogenesis, accelerated gluconeogenesis, and decelerated glycogenolysis. Conversely, overexpression of G0S2 inhibited fatty acid oxidation in mouse primary hepatocytes and caused sustained steatosis in liver accompanied by deficient TG clearance during the fasting-refeeding transition. In response to HFD, there was a profound increase in hepatic G0S2 expression in the fed state. Global and hepatic ablation of G0S2 both led to improved insulin sensitivity in HFD-fed mice. Our findings implicate a physiological role for G0S2 in the control of adaptive energy response to fasting and as a contributor to obesity-associated liver steatosis.     

Evidence for A1 and A3 receptors mediating adenosine-induced intracellular calcium release in the dorsal root ganglion neurons by using confocal microscopy imaging

Adenosine exerts a key role in analgesia. In the present study, adenosine-induced Ca²⁺ responses were revealed by using confocal microscopy imaging in the rat dorsal root ganglia (DRG) neurons in vitro. Our results showed that adenosine could evoke increases in the intracellular Ca²⁺ concentration in the DRG neurons. In addition, by application of selective receptor antagonists, two types of receptors, A₁R and A₃R, were identified to be involved in the adenosine-induced Ca²⁺ release from intracellular stores in neurons. Altogether, these results suggest that confocal microscopy imaging combined with fluorescent dyes could help to detect the analgesic-induced ion signaling in single cell.     

A retrospective analysis of pathological changes of testicular tissue in normal adult rats

Rat testicular model is widely used in experiments on andrology, pharmacology and reproductive toxicology. Generally, normal adult rat is considered to have normal testes. However, whether normal adult rats appeared abnormal testes have not been evaluated. The objective of this study was to evaluate the incidence of abnormal testes in normal adult Sprague Dawley (SD) rats and pathological changes in testicular tissues. Six hundred and sixteen adult male SD rats used in previous studies as controls were retrospectively analysed. Testicular tissues were stained with haematoxylin–eosin for observation of pathology. Among 616 rats, 14 rats had pathological testes, and the incidence of abnormal testis was 2.3%. In the 14 rats with abnormal testes, 10 rats were microrchidia (71.4%) and four rats showed normal testicular size. Testicular abnormality included complete interruption of spermatogenesis, partial germ cell arrest, progressive hypospermatogenesis, seminiferous epithelia vacuolation and inflammatory status. Bilateral testicular tissues had similar pathological changes in abnormal testes. The findings in this study demonstrate that the normal adult rats have abnormal testes. We should pay attention to the possibility of abnormal testes when using normal adult male rats for establishing a testicular model.     

shRNA-mediated GSTP1 gene silencing enhances androgen-independent cell line DU145 chemosensitivity

Purpose

Design short hairpin RNA (shRNA) interference sequence to silence glutathione S-transferase P1 (GSTP1) gene of androgen-independent prostate cancer cell line DU145, and then to explore its effect on sensitivity to chemotherapeutics.

Methods

Target sequence was picked up to form the shRNA. DU145 cell was divided into five groups according to the shRNA added for transfection: shRNA255, shRNA554, shRNA593, negative-shRNA and blank group. Fluorescence microscope was used to pick up the shRNA with the highest transfection ratio. Western blotting and RT-PCR were taken to pick up the shRNA with the best gene silencing result. 3-(4,5-Dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide assay and terminal de-oxynucleotidyl transferase-mediated dUTP nick end-labeling assay were used to detect survival ratio and apoptosis ratio of DU145 administered of fluorouracil (5-FU) or paclitaxel (PA) at different concentrations before and after shRNA transfection.

Results

Three different shRNA oligonucleotides (shRNA255; shRNA554; shRNA593) targeting the coding sequence of GSTP1 mRNA and one negative control shRNA were constructed. The transfection ratio of shRNA554 (76.2 ± 0.68 %) was higher than that of shRNA255 (63.3 ± 1.04 %) (P < 0.01) or shRNA593 (72.7 ± 0.33 %) (P < 0.01). After transfection of shRNA554, the mRNA and protein of level were the lowest, P < 0.01. The survival ratio of DU145 administered with 5-FU of different concentrations (30, 60, 120, 240 μg/ml) declined after transfection (P < 0.01). Besides, the apoptosis ratio increased after transfection (P < 0.01). Similarly the survival ratio of DU145 administered with PA of different concentrations (0.2, 2, 10, 20 μg/ml) declined (P < 0.01) and the apoptosis ratio increased (P < 0.01) after transfection.

Conclusions

The gene GSTP1 silence via shRNA transfection to androgen-independent prostate cancer cell line DU145 enhances the sensitivity to chemotherapeutics.