Heterotopic Auxiliary Rat Liver Transplantation With Flow-regulated Portal Vein Arterialization in Acute Hepatic Failure

In acute hepatic failure auxiliary liver transplantation is an interesting alternative approach. The aim is to provide a temporary support until the failing native liver has regenerated.^1-3 The APOLT-method, the orthotopic implantation of auxiliary segments- averts most of the technical problems. However this method necessitates extensive resections of both the native liver and the graft.⁴ In 1998, Erhard developed the heterotopic auxiliary liver transplantation (HALT) utilizing portal vein arterialization (PVA) (Figure 1). This technique showed promising initial clinical results.^5-6 We developed a HALT-technique with flow-regulated PVA in the rat to examine the influence of flow-regulated PVA on graft morphology and function (Figure 2).A liver graft reduced to 30 % of its original size, was heterotopically implanted in the right renal region of the recipient after explantation of the right kidney.  The infra-hepatic caval vein of the graft was anastomosed with the infrahepatic caval vein of the recipient. The arterialization of the donor’s portal vein was carried out via the recipient’s right renal artery with the stent technique. The blood-flow regulation of the arterialized portal vein was achieved with the use of a stent with an internal diameter of 0.3 mm. The celiac trunk of the graft was end-to-side anastomosed with the recipient’s aorta and the bile duct was implanted into the duodenum. A subtotal resection of the native liver was performed to induce acute hepatic failure. ⁷In this manner 112 transplantations were performed. The perioperative survival rate was 90% and the 6-week survival rate was 80%. Six weeks after operation, the native liver regenerated, showing an increase in weight from 2.3±0.8 g to 9.8±1 g. At this time, the graft’s weight decreased from 3.3±0.8 g to 2.3±0.8 g.We were able to obtain promising long-term results in terms of graft morphology and function. HALT with flow-regulated PVA reliably bridges acute hepatic failure until the native liver regenerates.     

Transient Expression of Proteins by Hydrodynamic Gene Delivery in Mice

Efficient expression of transgenes in vivo is of critical importance in studying gene function and developing treatments for diseases. Over the past years, hydrodynamic gene delivery (HGD) has emerged as a simple, fast, safe and effective method for delivering transgenes into rodents. This technique relies on the force generated by the rapid injection of a large volume of physiological solution to increase the permeability of cell membranes of perfused organs and thus deliver DNA into cells. One of the main advantages of HGD is the ability to introduce transgenes into mammalian cells using naked plasmid DNA (pDNA). Introducing an exogenous gene using a plasmid is minimally laborious, highly efficient and, contrary to viral carriers, remarkably safe. HGD was initially used to deliver genes into mice, it is now used to deliver a wide range of substances, including oligonucleotides, artificial chromosomes, RNA, proteins and small molecules into mice, rats and, to a limited degree, other animals. This protocol describes HGD in mice and focuses on three key aspects of the method that are critical to performing the procedure successfully: correct insertion of the needle into the vein, the volume of injection and the speed of delivery. Examples are given to show the application of this method to the transient expression of two genes that encode secreted, primate-specific proteins, apolipoprotein L-I (APOL-I) and haptoglobin-related protein (HPR).     

Enteric release of vasoactive intestinal peptide after a peptone meal in the dog

Vasoactive intestinal peptide (VIP) concentrations were measured by radioimmunoassay in plasma from portal and peripheral venous blood obtained from six alert, non-anesthetized dogs before and after gastric infusion of a 10% peptone meal. Mean basal portal and cephalic vein plasma VIP concentrations were $\text{42 ± 11.7}$ and $\text{42 ± 8.0}\text{(S.E.M.) pg/ml}$, respectively. No significant changes in peripheral venous plasma VIP concentrations were noted after the peptone meal throughout the duration of the collection period. In contrast, however, the mean VIP concentration in portal plasma increased promptly after the peptone meal with a peak of $\text{79 ± 8.2}\text{pg/ml}$ ($\text{P < 0.02}$) occurring 8 min after infusion of the meal. This was followed by a gradual decline in portal plasma VIP levels, with a return to prefeeding concentrations at 60 min ($\text{44 ± 6.3}\text{pg/ml}$). Results of these studies demonstrate that following gastric infusion of a peptone meal in the dog, portal, but not peripheral, plasma VIP concentrations increase significantly. Failure to detect augmentation of peripheral vein VIP levels after the meal is probably due to hepatic clearance of VIP.     

门静脉压力测定方法研究现况

肝硬化失代偿期出现的门脉高压症严重威胁着患者的生命,门静脉压力测定是研究门脉高压症的必要手段,也是治疗效果判断的重要指标。在目前的临床和科研工作中测压的方法种类繁多,大体上可分为间接和直接测压两大类。由于不同方法受到各自干扰因素的影响,所以所测结果差异明显,变异较大,给实际的工作中带来诸多不便,因此发现一种安全、准确、简单的门静脉压力测定方法显得尤为重要。本文就目前国内外各种门静脉压力测定方法做一综述。     

Expression of calcium-binding protein regucalcin mRNA in rat liver is stimulated by calcitonin: The hormonal effect is mediated through calcium

The involvement of a hypocalcemic hormone calcitonin (CT) in the expression of hepatic Ca²⁺-binding protein regucalcin mRNA was investigated. The change of regucalcin mRNA levels was analyzed by Northern blotting using liver regucalcin complementary DNA (0.9 kb). A single oral administration of calcium chloride (100 mg Ca/100 g body weight) to rats induced a remarkable increase in the serum calcium concentration and a corresponding elevation of the liver calcium content during 120 min after the administration. Thyroparathyroidectomy (TPTX) did not cause a significant increase in the liver calcium content after calcium administration. Hepatic regucalcin mRNA level was markedly elevated by calcium administration; the level was about 180% of controls at 60 min after the administration. This increase was completely abolished by TPTX. A single subcutaneous administration of CT (synthetic eel CT; 25–100 MRC mU/100 g) to TPTX rats received oral administration of calcium (100 mg/100 g) produced a remarkable increase in hepatic regucalcin mRNA levels; the level was about 280% of controls with the dose of 25 MRC mU CT/100 g. The present finding suggests that the expression of hepatic mRNA is stimulated by CT, and that the hormonal effect is mediated through Ca²⁺ in rat liver.     

Polymer-based delivering of shRNA to rabbit aortic smooth muscle cells suppressed the expression of IGF-1R in vitro and in vivo

Restenosis is one of clinical limitations for vein graft in coron- ary bypass graft. It has been proved that signal pathway IGF-1 and its receptor （IGF-1R） activated by hemodynamic mechanical stretch are responsible for the vascular smooth muscle cells proliferation in vein graft neointima formation. Unfortunately, there is no routinely successful method to resolve this problem. Gene delivering to vein graft possesses great therapeutic potential to prevent neointima formation. Polymer is one kind of nanoparticles, which can activate the process of endocytosis of cells. In this study, we evaluated the transfeetion efficiency and therapeutic potential of polymer- based transfection of plasmids expressing GFP and shRNAs targeting IGF-1R （pGFPshlGF-1Rs） to smooth muscle cells and rabbit external jugular vein graft. Results showed that polymer-based transfection provided high efficiency of trans- gene expression in smooth muscle cells in vitro. In vitro, IGF- 1R-specific shRNA transfected by polymer inhibited IGF-1R protein expression by 52 ±3.6%, when compared with mock transfected cells. In vivo delivering efficiency of pGFPshlGF-IR plasmid into the rabbit external jugular vein graft was significantly high in the polymer-based trans- fection group, when compared with negative control group. In vivo, polymer-based transfection IGF-1R-specific shRNA efficiently inhibited the expression of IGF-1R protein by 77 ± 3.6%, 65.6 ± 4.9%, and 76.7 ± 4.3% at 24, 48, and 72 h, respectively, when compared with negative control group. Our findings indicated that polymer-based transfec- tion may be a promising technique that allows the targeting of gene therapy for vein graft restenosis.     

降钙素基因相关肽基因直接注射大鼠

所设计的基因表达一个含降钙素基因相关肽(CGRP,37个氨基酸)、及其N端信号肽和C端酰胺化位点的多肽.该基因克隆到哺乳动物表达载体pBPV上并构成pBPV-CGRP.将此质粒DNA直接注射自发高血压大鼠并导致其血压在注射样品后3～22 d内明显下降,下降幅度超过了12％,在第16天甚至达到了24.42％.基因直接注射比CGRP本身的降压作用维持时间更久.     

选择性入肝血流阻断应用于肝癌合并门脉高压手术中的疗效观察

目的：探究选择性入肝血流阻断（SPVE 法）在肝癌合并门脉高压手术中的临床应用效果。方法：选择我院2009 年10 月~2014 年10 月期间确诊为肝癌合并门脉高压的患者80 例,按照随机数字数表法分为观察组和对照组各40 例,观察组行SPVE法进行血流阻断,对照组行全入肝血流阻断法（Pringle 法）进行血流阻断。对比两组手术时间、阻断血流时间、手术中出血量、输血量、手术前后患者肝功能相关指标及术后并发症发生率。结果：两组手术时间和血流阻断时间对比差异无统计学意义（P>0.05）,而观察组术中出血量及输血量均小于对照组,差异具有统计学意义（P<0.05）;术后观察组血液中谷丙转氨酶（ALT）、谷草转氨酶（AST）和总胆红素（TB）水平均低于对照组,而血清蛋白（ALB）和血红蛋白（Hb）水平高于对照组,差异均有统计学意义（均P<0.05）;观察组并发症发生率为22.5%,明显低于对照组的37.5%,差异有统计学意义（P<0.05）。结论：SPVE法应用在肝癌合并门脉高压手术中,可以显著减少术中的出血量和输血量,有利于术后肝功能的恢复,有效地降低术后并发症的发生。     

外源性TH基因在帕金森氏病鼠脑内的表达——行为和TH免疫组化的研究

用成年大鼠７５只,给右侧黑质区注射６－羟基多巴胺（６－ＯＨＤＡ）,损毁黑质多巴胺能神经元,制备偏侧帕金森氏病（ＰＤ）鼠模型。四周后,注射阿朴吗啡（ＡＰＯ）诱发大鼠向左侧旋转。旋转数为每分钟７次以上的３５只ＰＤ鼠作实验用。其中实验组１５只,对照组２０只。向实验组ＰＤ鼠右侧纹状体多点植入含大鼠酪氨酸羟化酶ｃＤＮＡ（ＴＨｃＤＮＡ）的真核表达载体ｐＳＶＫ３－ＴＨ和脂质体Ｌｉｐｏｆｅｃｔｉｎ混合的基因转染复     

Dielectric Properties of Water-liquid Paraffin Emulsion Systems at the Microwave Frequency of 9.4 GHz

The state of water contained in emulsions, particularly in o/w emulsions, was studied as a model of water orientation at the peripheries of biomembranes. Dielectric measurements made at microwave frequency on emulsions containing water and liquid paraffin in various ratios with emulsifiers revealed that the o/w emulsions possessed considerably reduced dielectric loss as compared with theoretical values obtained in accordance with the Maxwell-Wagner model, while the dielectric properties of w/o emulsion were in good agreement with the theoretically expected values. The observations seem to be explained by assuming changes in the state of water in the oil-water interfacial layer in o/w emulsions. The preparation of stable emulsions for use in this study is also discussed.