肝硬化大鼠肝缺血再灌注损伤肝细胞死亡方式

目的研究肝硬化大鼠肝缺血再灌注(I/R)损伤时肝细胞死亡的主要方式。方法采用四氯化碳复合法制作肝硬化大鼠模型,将肝硬化大鼠随机分为假手术(SO)组和I/R组,I/R组建立70%肝I/R模型,并于再灌注后0、1、6、24、48 h取材。比较各组的血清丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)水平,肝细胞钠钾ATP酶、钙ATP酶活性,流式细胞仪检测肝细胞胀亡和凋亡百分比,电镜观察肝细胞形态学改变。结果与SO组相比,I/R组再灌注后血清ALT、AST水平显著升高(P<0.05),6 h达高峰,后逐渐下降;肝细胞钠钾ATP酶、钙ATP酶活性显著下降(P<0.05),于再灌注1 h达低谷,后逐渐恢复;再灌注早期(6 h内)肝细胞死亡方式以胀亡为主,后期(24 h后)凋亡细胞逐渐增多;电镜下可见典型胀亡和凋亡改变。结论胀亡是肝硬化大鼠肝脏I/R损伤肝细胞死亡的主要方式,肝功能损伤与胀亡密切相关。     

多肿瘤标志物C12蛋白芯片对晚期胃肠癌的诊断有一定价值

目的:探讨多肿瘤标志物C12蛋白芯片检测系统在胃肠癌的诊断价值。方法:分析总结329例胃肠癌初治患者C12芯片的检测结果,寻找出与胃肠癌相关性最强的肿瘤标志物,计算各种肿瘤标志物组合方式对提高诊断率的贡献。结果:C12系统对本组329例胃肠癌患者的总体诊断率为39.21%,Ⅰ、Ⅱ、Ⅲ、Ⅳ期患者的诊断率分别为13.73%、33.33%、38.30%、58.03%。肿瘤标志物阳性率在各临床分期中的整体差异具有统计学显著性(P<0.01),Ⅰ期与Ⅲ期、Ⅰ期与Ⅳ期、Ⅱ期与Ⅳ期差异具有统计学显著性(P<0.01),其余分期相比没有统计学显著性(P>0.05)。C12系统中阳性率最高的三种TM是CEA、CA242和CA19-9,阳性率分别为27.36%、19.76%和19.45%,并且与肿瘤的分期相关。结论:C12检测系统对晚期胃肠癌的诊断有一定的价值,但对早期的敏感性不高,临床迫切需要微型高效的胃肠癌检测系统。     

Ferumoxide标记Flk1＋＋CD31＋-CD34-人骨髓间充质干细胞及其在食蟹猴脑内移植示踪观察

目的探讨Ferumoxide-PLL标记Flk1 CD31-CD34-人骨髓间充质干细胞(hBMSC)的方法及其在食蟹猴脑实质内移植活体示踪的可行性。方法采用Ferumoxide-PLL标记hBMSC,台盼蓝染色、普鲁士蓝染色和透射电镜扫描鉴定标记效率及细胞活力。体外磁共振成像(MRI)分别扫描标记和未标记细胞,计算T2*的弛豫时间和弛豫率(R2*)变化。通过立体定向手术将标记的hBMSC移植入食蟹猴右侧基底节区,采用MRI扫描活体示踪细胞。采用免疫组织化学、普鲁士蓝和HE染色对脑组织切片进行干细胞存活、分化及病理学研究。结果Ferumoxide-PLL标记hBMSC效率为96%,普鲁士蓝染色、电镜可显示标记hBMSC细胞质内铁颗粒。1×106和5×105两组Ferumoxide-PLL标记细胞的T2*的弛豫时间分别为68.86和79.88ms,而未标记细胞分别为12.71和15.24ms。标记细胞的R2*分别为78.68和65.61/s,分别是未标记细胞(14.52和12.52/s)的5.4和5.2倍。移植后3周MRI扫描T2WI仍可发现hBMSC呈明显的低信号。病理及免疫荧光结果显示hBMSCs在移植区大量存活,移植区有大量新生血管,但未见hBMSC向神经细胞分化。结论Ferumoxide-PLL可高效标记hBMSC,能显著增加其MRI图像对比度。MRI可活体示踪干细胞。移植入食蟹猴脑内的hBMSC可大量存活并促进新生血管形成。     

Blood pressure change during phacoemulsification and femtosecond laser-assisted cataract surgery

AIM: To evaluate blood pressure (BP) changes during phacoemulsification (PC) and femtosecond laser (FSL)-assisted cataract surgery. METHODS: A retrospective chart review was performed for all patients who received traditional phacoemulsification surgery (PC group) and FSL-assisted cataract surgery (FS group) from July 2013 to December 2014. Totally 206 eyes from 133 patients receiving the two types of procedures were included. Patient characteristics (age, gender, and hypertension history), pre- and post-operative BPs were collected. RESULTS: The pro-operative systolic and diastolic BPs (mm Hg) were 124.89±20.48 vs 126.98±16.85, and 71.88±9.81 vs 73.56±10.03, in PC and FS groups, respectively. While the post-operative systolic and diastolic BPs (mm Hg) were 130.13±22.59 vs 134.77±17.52, and 73.41±11.62 vs 78.89±12.2, in PC and FS groups, respectively. Paired-sample t-tests showed obvious systolic and diastolic BP elevations in FS group after surgery (P=0.001 and 0.007) and no reliability in PC group (P=0.094 and 0.359). A linear regression model revealed systolic and diastolic BP elevations, which were related to longer surgical times for FS group (P=0.008 and 0.021). Age, gender, and hypertension history were not correlated with blood pressure elevation in either group. CONCLUSION: BP increases but at a limited level after FSL-assisted cataract surgery compared to traditional phacoemulsificationKEYWORDS: cataract surgery; blood pressure; femtosecond-assisted cataract surgery; phacoemulsification     

Ligation of lymphocyte function-associated antigen-1 on monocytes decreases very late antigen-4-mediated adhesion through a reactive oxygen species-dependent pathway

Monocyte-endothelial adhesion plays an important role in monocyte trafficking and hence is important for immune responses and pathogenesis of inflammatory diseases including atherosclerosis. The cross-talk between different integrins on monocytes may be crucial for a coordinated regulation of the cellular adhesion during the complex process of transendothelial migration. By using monoclonal antibodies and recombinant intercellular adhesion molecule 1 (ICAM-1) to engage lymphocyte function-associated antigen 1 (LFA-1) on monocytic cells, we found that the cellular adhesion to vascular cell adhesion molecule 1 (VCAM-1) mediated by very late antigen 4 (VLA-4) was suppressed after this treatment and the suppression depended on the presence of reactive oxygen species (ROSs). Inhibition of production of ROSs through the use of inhibitor of the nicotinamide adenine dinucleotide phosphate (NADPH) oxidase, but not inhibitors of mitochondrial electron transport chain or xanthine oxidase, revealed that this suppression on VLA-4-mediated cellular binding was mediated by ROSs produced by phagocyte NADPH oxidase. Activation of phosphoinositol-3 kinase and Akt appears to mediate this NADPH oxidase activation through p47phox phosphorylation and Rac-1 activation. Our results provide a novel pathway in which ROSs play a critical role in integrin cross-talk in monocytes. This signaling pathway may be important for cellular transition from firm arrest to diapedesis during monocyte trafficking.     

In vivo and in vitro inhibitory effects of a traditional Chinese formulation on LPS-stimulated leukocyte-endothelial cell adhesion and VCAM-1 gene expression

Aim of the study

A traditional Chinese formulation Huang-Lian-Jie-Du-Tang (HLJDT) exerts anti-inflammatory effects. The present study aimed to investigate the effect of HLJDT on the LPS-stimulated leukocyte–endothelial cell adhesion and VCAM-1 gene expression both in vivo and in vitro.

Materials and methods

HLJDT was extracted from rhizoma coptidis, radix scutellariae, cortex phellodendri and fructus gardeniae in a weight rario of 1:1:1:1. In vivo leukocyte–endothelial cell adhesion was observed in rat lung after LPS stimulation (5 mg/kg, i.p.) with or without HLJDT (350 or 700 mg/kg, i.g.) pretreatment. The protein expression of vascular cell adhesion molecule 1 (VCAM-1) was analyzed by immunohistochemical method. In vitro leukocyte–endothelial cell adhesion was performed by examining the adhesion of THP-1 cells to LPS-stimulated human vascular endothelial cells with or without HLJDT pretreatment. The VCAM-1 expression at the RNA and protein levels was investigated by RT-PCR and western blot analysis, respectively. The activation of NF-κB was examined by the nuclear translocation of NF-κB by immunocytochemical method.

Results

In vivo, HLJDT dose-dependently reduced the number of leukocytes adhered to endothelium and VCAM-1 protein expression in lung venules of LPS-challenged rats. In vitro, HLJDT dose-dependently decreased the number of THP-1 cells adhered to LPS-stimulated endothelial cells and the expression of VCAM-1 both at the RNA and protein levels. The LPS-induced nuclear translocation of NF-kappa B in endothelial cells was also dose-dependently inhibited by HLJDT.

Conclusions

The present study demonstrated an additional mechanism underlying the anti-inflmmatory effect of HLJDT by inhibiting the leukocyte–endothelial cell adhesion and VCAM-1 gene expression. The inhibition of NF-kappa B activation by HLJDT might suggest a profound anti-inflammatory consequences.     

A community-based study of the relationship between insulin resistance/beta-cell dysfunction and diabetic retinopathy among type II diabetics in Kinmen, Taiwan

PURPOSE: To explore whether insulin resistance and beta-cell dysfunction are both related to diabetic retinopathy (DR) in type II diabetics by using a community-based study in Kinmen, Taiwan. METHODS: A screening program for DR was performed by a panel of ophthalmologists who used ophthalmoscopy and 45 degrees color retinal photographs on dilated pupils to determine a consensus grade of diabetic retinopathy. Screening, which was conducted between 1999 and 2002, involved 971 patients diagnosed with type II diabetes. The Homeostatis Model Assessment (HOMA) method was used to determine insulin resistance and beta-cell dysfunction. RESULTS: Seven hundred twenty-five diabetics who attended ophthalmological fundus checkups were studied. The overall response rate was 75%. After excluding 10 insulin-treated diabetics, diabetic retinopathy at first eye screening among the remaining 715 diabetics was 18.5%. Based on the multiple logistic regression, DR was found to be strongly related to both baseline insulin resistance (IR) and beta-cell dysfunction regardless of duration of diabetes. The strength of the relationships was maintained after adjustment for confounders. Those who were in the 2nd, 3rd, and 4th quartile of HOMA IR had 1.38 times (95% CI: 0.62-3.05), 2.37 times (95% CI: 1.19-4.69), and 4.16 times (95% CI: 2.15-8.06) the risk for DR compared to that in the 1st quartile, respectively. A reduced risk for DR in relation to HOMA beta-cell dysfunction for the 2nd, 3rd, and 4th quartile were 64% (95% CI: 27%-82%), 82% (95% CI: 58%-92%), and 82% (95% CI: 60%-92%) compared to that in the 1st quartile, respectively. CONCLUSIONS: Insulin resistance and beta-cell dysfunction are both associated with diabetic retinopathy in type II diabetes.     

Safety and efficacy of topical anesthesia combined with a lower concentration of intracameral lidocaine in phacoemulsification: paired human eye study

PURPOSE: To assess the safety and efficacy of phacoemulsification under a topical anesthesia combined with intracameral lidocaine 0.5%. SETTING: Department of Ophthalmology, Chang Gung Memorial Hospital, Taoyuan, Taiwan, China. METHODS: A prospective randomized double-blind study was designed in which patients had phacoemulsification performed under topical anesthesia (4 drops of nonpreserved lidocaine 2%) with 0.15 mL intracameral placebo (balanced salt solution) in 1 eye (Group 1) and topical anesthesia with intracameral nonpreserved lidocaine 0.5% in the other eye (Group 2). Endothelial changes, including cell density, coefficient variation of cell size, and percentage of hexagonal cells, were measured by noncontact specular microscopy. Preoperative and postoperative best corrected visual acuity was also documented. The degree of pain throughout surgery was ranked on a 10-point visual analog pain scale. RESULTS: Thirty-three patients were recruited. There was no significant difference in preoperative and postoperative mean endothelial parameters between the 2 groups. Furthermore, mean endothelial cell loss was similar. Mild or no pain (score 0 to 1) was reported by 48.5% in Group 1 and 90.9% in Group 2. Patients reported less pain with combined topical and intracameral lidocaine anesthesia (P = .001, Mann-Whitney test). Vision was significantly improved in both groups. However, 1 patient in Group A developed vitreous loss as a result of involuntary eye movement. CONCLUSION: Combining topical anesthesia with intracameral lidocaine 0.5% [corrected] anesthesia was safe and effective in phacoemulsification with intraocular lens implantation.     

肝脏及肢体缺血预处理抗大鼠肝缺血再灌注损伤的实验研究

目的研究肝脏缺血预处理(经典缺血预处理IPC)的第一保护窗(FW)与肢体缺血预处理(远端缺血预处理RPC)的第二保护窗(SW)及两者联合应用对大鼠肝脏缺血再灌注(I/R)损伤的保护作用及可能机制。方法大鼠随机分成5组:I/R组不行预处理;IPC组以肝缺血5 min行预处理;RPC组以双后肢缺血5 min,反复3次行预处理;RPC+IPC组先行RPC,24 h后行IPC作预处理;S组仅行开腹,不行其他处理。3个预处理组及I/R组均行肝缺血1 h再灌注3 h。取血用于血清谷丙转氨酶(ALT)与血清谷草转氨酶(AST)检测。切取肝组织用于测定肿瘤坏死因子α(TNF-α)和热休克蛋白70(HSP70)的表达、湿干比(W/D)及观察显微、超微结构的变化。结果与I/R组比较,IPC组,RPC组及RPC+IPC组ALT,AST,W/D及TNF-α阳性表达均明显降低(P0.01),HSP70表达量明显增加(P0.01),肝脏的显微及超微结构损伤减轻;IPC,RPC,RPC+IPC组3组间各项指标差异无统计学意义(P0.05)。结论IPC的FW,RPC的SW及两者联合应用对大鼠肝脏I/R损伤均有明显的保护作用,三者在保护强度上无明显差异。其机制可能与抑制TNF-α的产生、诱导保护性蛋白HSP70的表达、减轻肝脏水肿、改善肝组织微循环有关。