Technique of Porcine Liver Procurement and Orthotopic Transplantation using an Active Porto-Caval Shunt

The success of liver transplantation has resulted in a dramatic organ shortage. Each year, a considerable number of patients on the liver transplantation waiting list die without receiving an organ transplant or are delisted due to disease progression. Even after a successful transplantation, rejection and side effects of immunosuppression remain major concerns for graft survival and patient morbidity. Experimental animal research has been essential to the success of liver transplantation and still plays a pivotal role in the development of clinical transplantation practice. In particular, the porcine orthotopic liver transplantation model (OLTx) is optimal for clinically oriented research for its close resemblance to human size, anatomy, and physiology. Decompression of intestinal congestion during the anhepatic phase of porcine OLTx is important to guarantee reliable animal survival. The use of an active porto-caval-jugular shunt achieves excellent intestinal decompression. The system can be used for short-term as well as long-term survival experiments. The following protocol contains all technical information for a stable and reproducible liver transplantation model in pigs including post-operative animal care.     

The expense for one implantation of a banked bone allograft from a cadaveric donor and the issues affecting current advanced medical treatment in the Japanese orthopaedic field

Demand for banked bone allografts is increasing in Japan; however, there are too few bone banks and the bone bank network is not well-established. One reason for this was lack of funding for banks. Bone banks had to bear all material expenses of banked bone allografts themselves because this was not designated a covered expense. In December 2004, the Japanese government started a new “Advanced Medical Treatment” administration system which allowed an approved institution to charge the expense of authorized advanced medical treatments directly to patients. The treatment named “Cryopreserved allogenic bone and ligamentous tissue retrieved from cadaveric donor” was approved as an advanced medical treatment in March 2007. We present the calculation method and the expense per implantation of a banked bone allograft from a cadaveric donor under this treatment and raise issues which affect this advanced medical treatment and remain to be resolved in the Japanese orthopaedic field.     

DETs缓冲液对肠道微生物DNA的保存效果

方便高效的保存方法是开展动物肠道微生物研究的重要前提,用T-RFLP（末端限制性片段长度多态性）结合16S rDNA分析评估了野外采样常用的DETs（20%DMSO,0.25 M sodium-EDTA,100 mM Tris,pH 7.5,and NaCl to saturation）缓冲液对不同保存时间（24h,7天,30天,90天）粪便样品肠道微生物DNA的保存效果。结果表明,DETs保存在30天保存时间内微生物群落结构无显著变化,95.6%的T-RFs仍然存在,能效好的保护肠道微生物DNA。     

Browning of chitooligomers and their optimum preservation

Chitooligomers have attracted much interest due to their unique biological activities. However, chitooligomers easily turn brown during shelf life. The factors influencing the browning of chitooligomers were investigated. The results indicated that the browning was attributed to the structure change of chitooligomers. The water-solubility, thermal stability and moisture–adsorption of chitooligomers decreased with the increase of browning. The time, temperature, pH, moisture, oxygen and reductant all had effect on the browning of chitooligomers. The optimal preservation condition for chitooligomers should be at low temperature and humidity, at pH below 4 or above 10, and in absence of oxygen.     

无细胞百日咳攻击菌的制备及液氮保存方法分析

为了制备无细胞百日咳效价测定用攻击菌并用液氮进行保存,对此方法进行评价。采用原代攻击菌复苏传代后测定细菌浓度,并用蛋白胨水稀释为27亿个菌/m l,分装冻存管中。放入液氮罐内,并对液氮保存的攻击菌进行相应的检测,用统计软件计算。结果显示,此方法制备的攻击菌与传统的攻击菌无显著性差异,冻后虽攻击菌的毒力有所下降,但都在100-1000/MLD的正常范围内,且冻后0、3、6、9、12月无显著性差异。同批支间差异小,CV<5%。通过此方法制备的攻击菌稳定性好,试验结果符合《中华人民共和国药典》要求,可用于无细胞百日咳的效价测定。     

几种真姬菇菌种保藏方法的保藏效果对比

【背景】菌种保藏方法与保藏效果是菌种质量的前提和保证。随着真姬菇Hypsizygus marmoreus栽培规模和产量的逐年递增,选择最佳的菌种保藏方法也越来越迫切。【目的】对真姬菇菌种的常见保藏方法及保藏效果进行分析和研究,寻找简便、高效、廉价的真姬菇菌种保藏方法。【方法】通过对比菌丝生长率、菌丝脱氢酶活性以及菌丝对色度培养基的脱色率等指标参数来评价保藏方法的保藏效果。【结果】通过对三大类27种真姬菇菌种保藏方法 3个月保藏效果的对比分析发现,三大类菌种保藏方法中保藏效果最好的是水溶液保藏法,其中以0.1%PEG6000水溶液保藏效果最佳;其次为木屑保藏法,其中以盐水浸泡过的杨木木屑保藏效果最好;斜面保藏法效果最差。正常保藏实验的效果好于加速保藏实验。【结论】与当前真姬菇菌种保藏的常规方法相对比,本实验所采用的菌种保藏方法大大拓宽了菌种保藏方法的种类与范围,提高了菌种保藏的效果。     

Structure based virtual screening of MDPI database: Discovery of structurally diverse and novel DPP-IV inhibitors

Inhibition of dipeptidyl peptidase IV (DPP-IV) has been emerged as a promising approach for the treatment of type 2 diabetes (T2D). Structure based virtual screening (SBVS) of Molecular Diversity Preservation International (MDPI) database was performed using Glide and Gold against DPP-IV enzyme. Six promising hits were identified and tested for DPP-IV inhibition. Three compounds were found to be active at low micromolar concentration. The 3-(1-hydrazinyl-1-(phenylamino)ethyl)-4-hydroxy-1-methylquinolin-2(1H)-one (compound A) was found to be the most potent hit with an IC₅₀ of 0.73 μM. These three compounds (A, B and D) were then assessed for their glucose lowering effects in glucose fed hyperglycemic female Wistar rats. The glucose lowering effects of compounds also confirms their potential as anti-diabetic agents. The present study demonstrates a successful utilization of in silico SBVS tools in identification of novel and potential DPP-IV inhibitor.     

猴头菌单核菌丝厚垣孢子的观察

多次培养发现猴头菌单、双核菌丝均可形成厚垣孢子。厚垣孢子纺锤形,多端生、中生,大小6~88~10祄,其中单核厚垣孢子略小。试验表明,2~4℃猴头菌厚垣孢子的保藏期达7年以上;缺氧保藏效果好,30~52h萌发,萌发率32%~54%。     

子宫动脉化疗栓塞术治疗胎盘植入61 例的临床分析

目的:探讨子宫动脉化疗栓塞术治疗胎盘植入的临床疗效及预后。方法:以胎盘植入患者61例为研究对象,按治疗方式不同分为保守治疗组31例和化疗栓塞组30例。保守治疗组仅单纯口服米非司酮治疗,而化疗栓塞组采用子宫动脉化疗栓塞术进行治疗。比较两组患者的临床疗效、手术时间、出血量、输血量、住院时间、胎盘排出和月经复潮时间,术后并发症的发生情况,并随访术后1年患者的月经情况和妊娠情况。结果:经子宫动脉化疗栓塞术后29例成功止血,有1例未能止血而行子宫切除术。栓塞术后未出现器官局部缺血坏死、神经损伤等严重并发症,发热、下腹痛为常见并发症。而保守治疗组的31例患者中,因治疗失败导致切除子宫的7例,仅24例有效保留子宫,成功率仅77.4%,严重影响患者以后的生育能力。6例发生宫腔感染,4例发生宫腔粘连。子宫动脉化疗栓塞组β-HCG恢复正常时间、输血量、胎盘完全清除时间、月经异常的发生情况均优于保守治疗组,差异有统计学意义(P0.05)。对子宫动脉化疗栓塞术组患者进行为期1年的随访,14例患者在术后1个月后有胎盘组织自阴道排出,有11例患者2个月后发现胎盘残留,予以清宫术,除3例失访病例,其余患者在随访时间内恢复正常月经,并有2例再次妊娠者。结论:采用子宫动脉化疗栓塞术治疗胎盘植入,其术前准备时间和手术时间均短,出血控制迅速且并发症少,有助于保留患者的子宫,提高患者的生活质量。     

The influence of various storage conditions on cell viability in amniotic membrane

Up to now freeze-dried, gamma-sterilised or glycerol-preserved amniotic membranes (AMs) have widely been used in the field of ophthalmology and wound care (e.g. leg ulcers, burns). After some preservation processes in use, like freeze-drying or glycerol-preserving, the cells in the AM are no longer viable. Within this study we evaluated the influence of different short-term and long-term storage conditions on cell viability in AM. Therefore AMs from cesarean section placentae were washed and biopsied to evaluate the microbiological status and to determine the viability of the tissue. Additionally, viability under various storage conditions was examined by assessment of mitochondrial activity. Preservation included temperatures above and below 0°C as well as various media compositions. As expected, cell viability in amnion decreases during storage, in fact the effect was more pronounced when stored frozen, but the higher viability of amnion obtained by storage above 0°C with medium is associated with the limitation to a short period of storage of about 28 days. The evaluated preservation methods are the basis for future non-clinical in-vivo studies in which the possible benefit of amnion as a viable biomaterial in wound healing will be investigated. A part of this work was presented at the World Congress on Tissue Banking in Rio in May 2005 and was honoured by the Poster Award Commission.