Hydrodynamic liver gene transfer mechanism involves transient sinusoidal blood stasis and massive hepatocyte endocytic vesicles

The present study contributes to clarify the mechanism underlying the high efficacy of hepatocyte gene transfer mediated by hydrodynamic injection. Gene transfer experiments were performed employing the hAAT gene, and the efficacy and differential identification in mouse plasma of human transgene versus mouse gene was assessed by ELISA and proteomic procedures, respectively. By applying different experimental strategies such as cumulative dose-response efficacy, hemodynamic changes reflected by venous pressures, intravital microscopy, and morphological changes established by transmission electron microscopy, we found that: (a) cumulative multiple doses of transgene by hydrodynamic injection are efficient and well tolerated, resulting in therapeutic plasma levels of hAAT; (b) hydrodynamic injection mediates a transient inversion of intrahepatic blood flow, with circulatory stasis for a few minutes mainly in pericentral vein sinusoids; (c) transmission electron microscopy shows hydrodynamic injection to promote massive megafluid endocytic vesicles among hepatocytes around the central vein but not in hepatocytes around the periportal vein. We suggest that the mechanism of hydrodynamic liver gene transfer involves transient inversion of intrahepatic flow, sinusoidal blood stasis, and massive fluid endocytic vesicles in pericentral vein hepatocytes.     

术中超声在脑肿瘤切除术中的应用价值

摘 要 目的：采用术中超声观察颅脑肿瘤的超声图像特点，探讨其在神经外科手术中的应用价值。材料与方法：2007年3月～2008年3月，11例颅脑肿瘤患者在手术中接受了术中超声检查，以确定肿瘤的位置，手术切除范围，及切除术后是否有肿瘤残余。结果：术中超声清晰的显示了肿瘤的部位，形态及回声，11例患者全部经手术完整切除肿瘤，均未损伤颅脑功能区。术后病理提示星形胶质细胞瘤5例、胶质增生1例、海绵状血管瘤2例、转移性肿瘤2例、恶性淋巴瘤1例。结论：术中超声作为一种实时、无创、简便的辅助检查方法，可帮助手术医师确定肿瘤的部位及边界，避开功能区，提高手术安全性及治疗效果。     

Efficacy of the potentized homeopathic drug, Carcinosin 200, fed alone and in combination with another drug, Chelidonium 200, in amelioration of p-dimethylaminoazobenzene-induced hepatocarcinogenesis in mice

OBJECTIVES: This study was conducted to examine whether the potentized homeopathic remedy Carcinosin 200, fed alone and in combination with Chelidonium 200, has differential protective effects against p-dimethylaminoazobenzene (p-DAB)-induced hepatocarcinogenesis in mice. DESIGN: Liver tumors were induced in mice through chronic feeding of p-DAB (initiator) and phenobarbital (PB, promoter). The mice were divided into two subgroups: (1) one was fed potentized Alcohol 200 and served as controls; and (2) the other was fed Carcinosin 200 alone or in combination with Chelidonium 200 and divided into several sets. The relative efficacy of the two potentized remedies, alone or in combination, in combating hepatocarcinogenesis was assessed through several cytogenetical endpoints such as chromosome aberrations, induction of micronuclei, sperm head anomaly, and mitotic index at several intervals of fixation (days 7, 15, 30, 60, 90, and 120). Several toxicity biomarkers such as acid and alkaline phosphatases, glutamate oxaloacetate transaminase, glutamate pyruvate transaminase, and lipid peroxidation activity were also assayed in three organs of treated and control mice. In addition, recovery by the homeopathic drugs, if any, of tissue damage inflicted because of chronic feeding of p-DAB and PB was also assessed by optical, scanning, and transmission electron microscopies of liver done at days 60 and 120. RESULTS: Both Carcinosin 200 and Chelidonium 200 when administered alone show considerable ameliorative effect against p-DAB-induced hepatocarcinogenesis in mice; but the conjoint feeding of these two drugs appears to have had a slightly greater protective effect. CONCLUSIONS: These homeopathic remedies have the potential to be used as complementary and alternative medicine in liver cancer therapy, particularly as supporting palliative measures.     

Inhibition of carrageenin-induced pedal oedema in rats by immobilisation stress

The effect of immobilisation stress on acute pedal inflammation induced by carrageenin, and the mechanism of stress-induced anti-inflammatory effect, were investigated in male Wistar strain albino rats. Carrageenin-induced pedal inflammation oedema was attenuated by immobilisation stress in a time-dependent manner, when the rats were restrained for 30 min, 1 h, and 2 h immediately after the induction of the inflammation. Pentobarbitone exhibited significant anti-inflammatory effect of its own in an anaesthetic dose and also inhibited stress (1 h)-induced attenuation of the inflammation. Likewise, lignocaine, injected behind the knee joint of the inflamed limb, attenuated the inflammation and also inhibited the stress-induced anti-inflammatory effect. These findings indicate the importance of the central nervous system (CNS) and the afferent/efferent neural pathways from and to the inflammatory site, in inflammation and in stress-induced anti-inflammatory effect. Earlier studies from this laboratory have shown that the central noradrenergic, histaminergic, serotonergic and GABA-ergic neurotransmitter systems have a modulatory anti-inflammatory effect on carrageenin-induced pedal oedema. Since all these neurotransmitter systems have been reported to be activated by stress, their role was assessed in the inflammation-attenuation effect of immobilisation stress. The present studies indicate that, of these neurotransmitters, only the central noradrenergic system is involved in the anti-oedema effect of stress. Endogenous opioid peptides may also be involved in the stress-inflammation interaction, since naloxone inhibited the stress effect. Bilateral adrenalectomy and peripheral chemical sympathectomy, induced by i.p. administration of 6-hydroxydopamine, augmented carrageenin oedema and antagonised the stress-induced anti-inflammatory effect. However, metyrapone, an inhibitor of endogenous corticoid synthesis, failed to inhibit the stress effect. These findings indicate that the sympatho-medullary system, which is known to be activated during stress, is responsible for the observed anti-inflammatory effect of immobilisation stress, rather than augmented release of adrenal corticoids. It is suggested that the observed inflammation reducing effect of immobilisation stress is a consequence of increased central noradrenergic and peripheral sympatho-medullary activity.