明胶降解物对大鼠真皮成纤维细胞增殖及Ⅰ型胶原蛋白分泌的影响

目的 探讨组织工程心瓣膜常用人工基质材料--明胶发生降解后的产物对大鼠真皮成纤维细胞增殖及Ⅰ型胶原蛋白分泌的影响. 方法 利用酶组合技术制备明胶降解物（gelatin hydrolysate, GH）,采用超滤的方法分离出低分子量明胶降解物组分（gelatin hydrolysate fraction, GHF）,用于5只SD大鼠真皮成纤维细胞的体外培养.采用CCK-8活细胞计数试剂盒（Cell Counting Kit-8）检测细胞的增殖情况,ELISA试剂盒检测细胞I型胶原蛋白的分泌情况. 结果 ①GHF对细胞增殖作用的浓度效应：在0.125～1.0 mg/ml浓度范围内,GHF对细胞的促增殖作用呈现先增强后减弱的趋势,在0.25 mg/ml时,细胞增殖率最大,达到（47.54±16.35）%;②GH与GHF对细胞增殖影响比较：GH与GHF均具有促进细胞增殖的作用,且在浓度为0.25 mg/ml时,促细胞增殖作用无显著差异[（27.04±15.21）% vs （39.22±15.55）%, P=0.177];③GH与GHF对细胞分泌Ⅰ型胶原蛋白影响比较：GHF组第3天时Ⅰ型胶原分泌量为（28.06±5.60）pg/105,与空白对照的（18.24±4.52）pg/105相比,有显著差异（P=0.006）,而GH组直至第6天时Ⅰ型胶原蛋白分泌量,与空白对照相比,无统计学意义（P=0.103）. 结论 分子量大小不同的明胶降解物GH与GHF均具有促进大鼠真皮成纤维细胞增殖的作用,且分子量较小的GHF还具有促进细胞I型胶原分泌的功能.     

脂膜微泡结合凝血酶原复合物的制备

目的:制备结合凝血酶原复合物(PCC)的阴离子脂膜微泡,评价微泡的理化性质。材料和方法:采用机械振荡直接连接法,分两组(于机械振荡之前或之后加入),制备结合PCC的阴离子脂膜微泡,观察并检测洗涤前后微泡的理化性质。结果:微泡与PCC的结合率及Ⅸ因子活性:两组间比较无明显差异(P>0.05),但洗涤后结合率由洗涤前的95%降为80%,活性由90%降为19%。结论:直接连接法可制备结合PCC的阴离子脂膜微泡,微泡与PCC结合率较高,且洗涤前能保持Ⅸ因子较高的活性。     

关节镜下人工韧带重建前交叉韧带的临床初步体会

目的探讨关节镜下应用LARS人工韧带重建前交叉韧带的可行性及近期疗效. 方法用法国产LARS人工韧带对16例前交叉韧带(anterior cruciate ligament,ACL)损伤行关节镜下ACL重建术.等距点钻胫骨、股骨骨道,将肌腱拉入骨道,韧带游离部分位于关节腔内,拉紧后2枚螺钉固定韧带,合并损伤同期处理. 结果手术时间51～86 min,平均64 min.术后无滑膜炎、韧带断裂、活动明显受限等并发症.16例均随访1.5～6个月,平均3.8月.按照IKDC评分标准:术前C级6例,D级10例;术后A级6例,B级9例, C级1例(χ2=6.264,P<0.05).Lysholm膝关节功能评分术前36～76分,(63.7±7.3)分;术后86～97分,(94.8±9.6)分(t=10.356,P<0.05). 结论关节镜下LARS人工韧带重建ACL,操作简便,可使膝关节获得即时稳定性,早期康复锻炼,最大限度的防止关节功能受限,近期疗效满意.     

Protein adsorption on ex vivo catheters and polymers exposed to peritoneal dialysis effluent.

BACKGROUND: Deposition of proteins on surfaces of medical devices has been recognized to putatively relate to the process of regulation of biomaterial-associated complications by attachment of fibrin clots, eukaryotic cells, and microbes. The molecules adsorb to a varying extent, depending not only on the physicochemical properties of the biomaterial, but also on the composition of the host fluid. OBJECTIVE: Adsorption of proteins on catheters exposed both ex vivo and in vitro to dialysate of patients on peritoneal dialysis (PD) was studied. METHODS: Peritoneal dialysis effluent was collected from 5 patients with end-stage renal disease on continuous ambulatory PD. Tenckhoff catheters were obtained from 16 patients. Deposition of proteins on excised Tenckhoff catheters and tubing of different materials exposed to PD effluent in vitro was studied using 125iodine-labeled antibodies. Adhesion of Staphylococcus aureus and Staphylococcus epidermidis strains was quantified on tubing exposed to PD effluent in vitro. RESULTS: The presence of albumin, transferrin, immunoglobulin G, fibrinogen, fibronectin, von Willebrand factor, vitronectin, and thrombospondin was determined at various concentrations in PD effluent. All proteins analyzed were detected on PD catheters removed from patients. The extent of protein deposition on Tenckhoff catheters exposed to PD effluent, in vitro, rapidly reached a plateau and remained constant, as it did on polyvinyl chloride and polyethylene tubing. Adhesion of staphylococci was enhanced on Tenckhoff catheters exposed to PD effluent compared to unused PD solution. CONCLUSIONS: The data identify surface exposed proteins that may serve as adhesion sites for microbes on peritoneal catheters indwelled in patients undergoing PD.     

Immunology and genetics

Medical and Surgical Dermatology -     

派罗欣治疗慢性乙型病毒性肝炎的观察与护理

目的探讨乳腺癌术后胸部气压固定带的制作及临床应用价值。方法将64例乳腺癌术后患者随机分为观察组和对照组各32例,对照组采用传统的绷带加压包扎,观察组采用自行设计并制作的胸部气压固定带加压包扎。观察两组术后引流量、拔管时间及并发症的发生情况。结果观察组皮下积液发生率显著低于对照组（P〈0.05）,术后24h引流量、置管时间显著少于/短于对照组（均P〈0.01）。结论乳腺癌术后使用胸部气压固定带加压固定可减少渗出,缩短置管时间,降低皮下积液发生率。     

MED术中应用自制器械处理后纵韧带钙化及骨赘增生

目的探讨脊柱后路显微内镜(microendoscopic disectomy,MED)手术治疗腰椎间盘突出症合并后纵韧带钙化及椎体后缘骨赘增生的方法. 方法 MED手术中应用自制弧形纤维环刀和L形打入器治疗115例该类患者. 结果全组随访12～30个月,平均22个月,按MacNab标准:优、良109例,可5例,差1例,优良率94.8(109/115).未出现硬脊膜破裂及神经孙上,未发生椎间隙感染,无中转开放手术. 结论正确运用MED技术,配合采用自制器械及相应的改良方法,能较好地处理合并后纵韧带钙化及椎体后缘骨赘增生的难题,进一步扩大了手术适应证范围,提高了疗效及安全性.     

BKV in Simultaneous Pancreas-Kidney Transplant Recipients: A Leading Cause of Renal Graft Loss in First 2 Years Post-Transplant

With the introduction of more potent immunosuppressive agents, rejection has decreased in simultaneous pancreas/kidney transplant (SPK) recipients. However, as a consequence, opportunistic infections have increased. The purpose of this report is to outline the course of SPK patients who developed polyomavirus-associated nephropathy (PVAN). A retrospective review of 146 consecutive SPK recipients from January 1, 1996 to December 31, 2002 was performed. Immunosuppression, rejection and development of PVAN were reviewed. Nine patients were identified. All received induction with either OKT3 or thymoglobulin. Immunosuppression included tacrolimus/cyclosporine, MMF/azathioprine and sirolimus/prednisone. Two patients were treated for kidney rejection prior to the diagnosis of PVAN. Time to diagnosis was an average of 359.3 days post-transplantation. Immunosuppression was decreased but five ultimately lost function. However, none developed pancreatic abnormalities as demonstrated by normal glucose and amylase. Two underwent renal retransplantation after PVAN diagnosis and both have normal kidney function. PVAN was the leading cause of renal loss in SPK patients in the first 2 years after transplantation and is a serious concern for SPK recipients. The pancreas, however, is spared from evidence of infection, and no pancreatic rejection occurred when immunosuppression was decreased.     

Intra-lesional injections of collagenase are ineffective in the treatment of keloid and hypertrophic scars.

The treatment of keloid and hypertrophic scars remains difficult. Enzymatic digestion of keloid scars has been previously proposed as an effective treatment strategy for reducing the volume of keloid scars. To test this, we administered intra-lesional injections of pure collagenase (between 600 and 4500 units for each scar) into the keloid and hypertrophic scars of seven human volunteers (five keloid and two hypertrophic scars). Five patients (three keloid and two hypertrophic) received more than one injection of collagenase. The treatment resulted in a temporary reduction in scar volume for three of the patients with keloid scars. However, scar volumes for these three patients returned to the same (or greater) levels after 6 months of follow-up. Treatment with collagenase produced no change in scar volume for the two patients with hypertrophic scar. Side effects were numerous and severe including; pain, swelling, blistering, ulceration and ecchymosis at the site of injection. One patient required admission to hospital for 48 h after the first injection. Maximum length of follow-up was 6 months. None of the seven patients completed the study and returned for final follow-up at 2 years. This pilot study suggests that treatment of keloid and hypertrophic scars with intra-lesional injections of collagenase is ineffective.     

Fabrication of porous hollow silica nanoparticles and their applications in drug release control.

Preparation and characterization of porous hollow silica nanoparticles (PHSN) for controlled release applications were investigated. Through orthogonally designed experiments, the optimal synthesis conditions for the preparation of PHSN were obtained and the produced PHSN were characterized by BET, SEM, TEM and IR. Scanning and transmission electron microscopy images revealed their hollow shell-core structure and also demonstrated that the size and shape of PHSN are determined by the templating CaCO3 nanoparticles. The produced PHSN were applied as a carrier to study the controlled release behaviors of Brilliant Blue F (BB), which was used as a model drug. Being loaded into the inner core and on the surfaces of the nanoparticles, BB was released slowly into a bulk solution for about 1140 min as compared to only 10 min for the normal SiO2 nanoparticles, thus exhibited a typical sustained release pattern without any burst effect. In addition, higher BET of the carriers, lower pH value and lower temperature prolonged BB release from PHSN, while stirring speed showed little influence on the release behavior. It showed that PHSN have a promising future in controlled drug delivery applications.