胃黏膜单细胞悬液的制备及其周期蛋白的表达

目的寻找一种方法能够有效去除胃黏液，把胃黏膜制备成单细胞悬液以进行胃黏膜流式细胞术等单细胞的蛋白分析研究，为即将在胃组织开展细胞组学做好准备。方法应用酶反应去除胃黏液和分离黏膜层后．再使用机械法制备成单细胞悬液，并应用细胞免疫组织化学流式细胞术和激光共聚焦显微镜技术初步检测胃黏膜细胞主要周期蛋白cyclin D、E、A、B1的表达。结果0．1％的胃蛋白酶和1．2-2．4U／L的中性蛋白酶分别能够较好地去除胃黏液，获得黏膜层，从而制备成胃黏膜单细胞悬液，从流式中首次系统地显示出其细胞主要周期蛋白的时相性表达规律：cyclin D3、B1明显表达，cyclin D2较弱，cyclin D1、A、E表达不明显，并得到激光扫描共聚焦显微镜的验证。结论利用胃蛋白酶，中性蛋白酶作用后能够去除胃黏液．较容易获得胃黏膜单细胞悬液；并提示体内的细胞周期蛋白的表达规律与体外培养的细胞系的表达规律不同。     

腹腔镜手术对腹腔内环境的影响

目的 研究腹腔镜手术对腹腔内的pH值、中性粒细胞吞噬功能及红细胞Ⅰ型补体受体(CR1)免疫粘附活性的影响。方法 对32例腹腔镜胆囊切除术患者术中留取腹腔内血性渗液测定其pH值、中性粒细胞吞噬功能、红细胞CR1免疫粘附活性，并与术中测定的周围血相应值比较。结果 腹腔镜手术时，血液中的pH值为7．4l，吞噬细胞百分数及吞噬指数分别为74．6％及55．7，红细胞CR1免疫粘附活性为58％；而腹腔渗液的pH值为7．13，吞噬细胞百分数及吞噬指数分别为65．6％及47．6，红细胞CR1免疫粘附活性为50％。结论 腹腔镜手术中CO2气腹造成腹腔内pH值明显下降，腹腔内中性粒细胞吞噬功能及红细胞CR1免疫粘附活性也明显降低。因此，严重炎症患者在腹腔镜手术时，在CO2气腹压力作用下，可能引起炎症扩散。     

白蛋白刺激肾小管上皮细胞表达基质金属蛋白酶2和9

目的探讨白蛋白对近端肾小管上皮细胞表达基质金属蛋白酶2（MMP-2）和MMP-9的影响。方法大鼠近端肾小管细胞株NRK52E培养至70％或100％融合时，分别给予不同浓度（0．1～1．0g／L）去脂和非去脂牛血清白蛋白（dBSA和BSA）刺激，于24、48、72h收集培养液，用明胶酶谱和Western印迹方法检测培养液MMP-2和MMP-9活性和蛋白水平。结果与空白对照组比较，1．0g／LBSA刺激未完全融合NRK52E72h后，MMP-2和MMP-9活性分别上调276％、176％（P〈0．05）。与刺激24h比较，1．0g／LBSA刺激72h，在未完全融合NRK52E的MMP-2和MMP-9活性分别上调536％、148％；在完全融合NRK52E分别上调212％、184％（P〈0．05）。与完全融合NRK52E比较，1．0g／LBSA刺激未完全融合NRK52E 72h，MMP-2活性上调增加了274％，MMP-9活性上调减少了45．1％（P〈0．05）。dBSA刺激结果与BSA类似。结论白蛋白刺激呈剂量和时间依赖性上调近端肾小管细胞表达MMP-2和MMP-9。细胞完全融合可抑制MMP-2表达，促进MMP-9表达。     

单纯疱疹病毒Ⅰ型扩增子载体转移脑啡肽基因在神经细胞的表达

目的：研究采用单纯疱疹病毒I型扩增子载体将脑啡肽基因导入神经细胞，观察载体lacZ基因和外源脑啡肽基因的表达。方法：实验共分4组；A组为对照组，未加病毒悬液的神经细胞；B组，加病毒悬液的神经细胞培养2d；C组，加病毒悬液的神经细胞培养3d；D组，加病毒悬液的神经细胞培养至第10d。留部基用放射免疫法检测外源性脑啡肽基因的表达。用X-gal原位检测lacZ基因表达反映扩增子病毒的存在和滴度。结果：L－脑啡肽放射免疫检测证实转基因神经细胞L－脑啡肽含量较对照组明显升高（P＜0．05），X-gal染色证实加入假病毒悬液的神经细胞有30％以上的细胞蓝染.结论：HSV－1扩增子假病毒能将外源脑啡肽基因导入神经细胞，并使之有效表达。     

含不同浓度参附注射液的停跳液对兔离体心脏缺血再灌注损伤的保护作用

目的探讨含不同浓度参附的停跳液对兔离体心脏缺血再灌注损伤的保护作用及其机制。方法40只成年大耳白兔，建立Langendorff模型灌注离体心脏，随机分为5组，每组8只。对照组：用St．Thomas停跳液使心脏停搏；SF1、SF5、SF10、SF15组：分别予含1％、5％、10％、15％参附注射液（SFI）的St．Thomas停跳液使心脏停搏。平衡灌注30min时各组分别给予不同停跳液20ml，停灌45min时再灌注40min，测定再灌注后冠脉流出液中磷酸肌酸激酶同工酶（CK-MB）活性，分离心肌线粒体，测量丙二醛（MDA）、Ca^2＋含量，电镜观察心肌超微结构，行线粒体Flameng评分，并用比表面（δ）和面数密度（NA）进行线粒体体视学评分。结果与对照组比较，SF1、SF5、SF10组冠脉流出液中CK-MB活性降低，线粒体MDA、Ca^2＋含量及Flameng评分降低，线粒体NA增高，SF5、SF10组线粒体δ增高。SF15组冠脉流出液CK-MB活性增高，线粒体MDA含量降低，Ca^2＋含量升高（P〈0．05或0．01）。SF5、SF10组心肌病理学损伤较对照组轻。结论含5％-10％SFI的St．Thomas停跳液对心肌线粒体有较好的保护作用。减轻心肌缺血再灌注损伤。     

柚子提取物对小鼠肝微粒体代谢活性的影响

目的探讨几种柚子提取物对肝微粒体代谢活性的抑制效应。方法在体外小鼠肝微粒体孵育体系中加入睾丸酮和不同柚子提取物后，以高效液相色谱法（HPLC）测定6-β羟基睾丸酮的生成量。结果6—β羟基睾丸酮的线性回归方程为y=0．0904x＋0．1795，r=0．9986，x表示峰面积，y表示6-β羟基睾丸酮的含量，单位nmol／ml。4种柚子提取物A皮、C皮、A肉、C肉对6-β羟基睾丸酮生成的抑制率分别为47．9％、28．8％、82．4％和72．3％。结论几种柚子提取物较对照组对肝微粒体的代谢活性均有抑制效应，其中A肉的抑制率最大。     

新型支架材料磷酸钙骨水泥-聚乳酸聚羟基乙酸二聚体复合物生物相容性的研究

目的 评价新型支架材料磷酸钙骨水泥-聚乳酸聚羟基乙酸二聚体（CPC-PLGA）复合物的生物相容性。方法 噻唑蓝（MTT）比色法分析3种浓度材料浸提液（100％、50％、10％）对L929细胞生长的影响；分光光度法分析3种浓度材料浸提液的溶血性；复合物植入SD大鼠臀肌2周和4周后取出评价组织相容性；种植BMSC1周扫描电镜观察生长状况。结果 24h3种浓度材料浸提液RGR分别为76％、76％、85％，48h分别为75％、84％、96％，72h分别为76％、77％、84％，细胞毒性评级均为1级；3种浓度浸提液对健康人血的溶血率分别为3．45％、2．59％、0．86％；肌肉内植入2周复合物组炎性反应为Ⅰ级4例、Ⅱ级5例、Ⅲ级1例，纤维囊腔形成10例均为Ⅳ级；4周复合物组炎性反应为Ⅰ级5例、Ⅱ级5例，纤维囊腔形成Ⅲ级3例、Ⅳ级7例；种植1周BMSC形态正常、生长状态良好。结论 CPC-PLGA复合物无细胞毒性，无溶血作用，植入后无炎症反应，种植细胞生长良好，具有良好的生物相容性。     

黑素细胞悬液中加入透明质酸的最佳浓度选择

目的：观察透明质酸对黑素细胞增殖和酪氨酸酶活性的影响，摸索最佳复合浓度。方法：在初步测试透明质酸保持黏稠度的可稀释倍数基础上，用不同浓度透明质酸作用于贴壁的黑素细胞，用MTT方法和酪氨酸活性测定分别检测黑素细胞生长和黑素合成能力的变化；在黑素细胞接种同时和接种后加入透明质酸，检测对黑素细胞生长的影响。结果：透明质酸在2～4ug／μl对贴壁的黑素细胞增殖有明显促进作用，终浓度低于0．5ug／μ1时作用不明显，浓度达8ug／μl时促增殖作用反而下降；透明质酸对黑素细胞酪氨酸酶活性影响结果类似。在接种同时加入透明质酸(1～4ug／μl)的黑素细胞增殖明显快于接种后加入透明质酸组。结论：透明质酸在2～4ug／μl浓度时能促进黑素细胞增殖，提高酪氨酸酶活性。用这种浓度的透明质酸混悬黑素细胞后可以增加细胞悬液的黏性，维持细胞在移植床上驻留，促进细胞增殖，有助于移植治疗白癜风。     

大鼠侧脑室注射细胞因子等对脑损伤的形态学研究

目的：探讨大鼠侧脑室注射细胞因子和高渗液等致病因素对脑组织的损害作用。方法：48只雄性SD大鼠分为6组：侧脑室内分别注射TNF—α、10％NaCl、胰脂肪酶、白色念珠菌悬液及大肠埃希菌悬液，注射0．9％NaCl作为对照；以光镜和电镜观察不同损害因素对大鼠脑组织的作用。结果：TNF—α、10％NaCl、胰脂肪酶、真菌及细菌悬液均可招致不同程度的脑神经元和胶质细胞的水肿、核固缩、脑软化、轴突髓鞘板层结构紊乱、髓鞘脱失等病理改变。结论：促炎细胞因子、高渗状态、胰脂肪酶以及真菌和细菌感染都是重症急性胰腺炎病程中导致脑组织髓鞘、神经元和胶质细胞等损伤的病理因素。     

甜菊甙研制中改进絮凝工艺的研究

作者研究了甜菊甙分离过程中的提取液和大孔径吸附树脂解吸液的絮凝工艺。对Ca(OH)_2调节的各种pH值条件下去杂效果及机理作了探讨,发现pH在7—8间各种非糖甙杂质除去率较高。在比较了各种絮凝剂的絮凝效果及脱色率后,发现聚氯化铝的絮凝效果及脱色最佳。     

Prognostic significance of serum albumin urea ratio in suture line leakage of the alimentary tract

Prognostic significance of postoperative changes in serum albumin and urea concentrations as well as their ratio was statistically evaluated in 73 patients with suture line leakage of the abdominal alimentary tract. Increase in serum urea was associated with poor prognosis regardless of the site of leakage. Although the serum albumin concentration was poorly correlated to prognosis, its ratio to serum urea was of more prognostic significance than the urea value alone. When the ratio of the serum albumin nitrogen to the urea nitrogen fell below 10, there was a sharp rise in mortality as well as in morbidity, associated with poor local defense reaction, high incidence of spreading peritonitis, severe systemic complications, and decreased tolerance to tube feeding as well as parenteral hyperalimentation. A special therapeutic policy was suggested for the management of leakage in patients with critically low albumin/urea ratios.     

Comparison of the stability of press-fit hip prosthesis femoral stems using a synthetic model femur

The motion of three different press-fit hip prosthesis femoral components was compared with that of a cemented stem and an Austin Moore noncemented hemiarthroplasty. Synthetic composite femurs were used as an experimental model to reduce the variations in shape and quality typical among cadaver femurs. Motion was measured under axial and rotational loading approximating a walking load. On the initial application of load, axial subsidence was as much as several millimeters for the noncemented stems. Considerable tightening occurred during the 5,000 walking cycles, such that the motion of the noncemented stems in some directions eventually was as small as that of the cemented stem, on the order of tens of microns.     

Preliminary investigations of spider silk in wounds in vivo — Implications for an innovative wound dressing

The ideal wound dressing in particular for burn wounds has not been found yet. The aim of this study was to investigate native spider silk as a novel wound dressing.Release of inflammatory cytokines of macrophages and neutrophile granulocytes was determined via ELISA after exposure to spider silk. Migration of dermal cells as well as angiogenesis on spider silk was visualized with live video microscopy or chorioallantois membrane model, respectively. Native spider silk was placed in full-thickness skin wounds in a sheep in vivo-model and wounds were evaluated after 2, 4, 6, and 8 weeks histologically as well as per quantitative real-time PCR.Minimal inflammatory cytokine release could be seen for spider silk. Ingrowth of single capillaries into bundles of spider silk and migration of keratinocytes as well as fibroblasts on spider silk fibres was proven. Macroscopically, a comparable wound closure could be seen in spider silk and in sham controls. In histological evaluation, a thicker epidermis was observed in spider silk treated wounds while collagen III/I expression ratio was comparable in both groups.As native spider silk has been described as highly biocompatible, it might represent an innovative alternative to common wound dressings.     

Spontaneous esophageal rupture treated with staged operations

Spontaneous esophageal rupture is a life-threatening entity. Here, a 64-year-old male who presented with sudden onset of severe back pain was diagnosed as having an esophageal rupture to the right pleural cavity. Emergency operation was carried out 16 hours after the onset. The rupture was as large as 7 cm and the surrounding tissue was fragile and necrotic. We performed an esophagectomy as a primary salvage procedure. An esophageal reconstruction was carried out successfully 6 months after the initial operation. Staged operative strategy including esophagectomy is still an important option to treat this kind of high risk patient.     

Vertical Banded Gastroplasty vs Adjustable Silicone Gastric Banding in the Treatment of Morbid Obesity: a Preliminary Report

A prospective comparative study, comparing vertical banded gastroplasty (VBG) with adjustable silicone gastric banding (ASGB) has been undertaken. The purpose of the study was to see if ASGB could produce weight loss as good as VBG. Patient population was similar and patients' choice was based on informed consent. Male/female ratio and excess body weight were comparable in both groups. Early as well as late complications were minor and rare in both groups. The evaluation of the results was based on excess weight loss and BMI curves. Weight loss as well as BMI curves were quite similar in the two groups. A 50% excess weight loss has been obtained at 6 months, in both groups.     

The Subchondral Bone of the Proximal Tibial Epiphysis in Osteoarthritis of the Knee

The trabecular bone of the proximal end of the tibia was assessed as an endoprosthesis-bearing structure. The mass and mineral content as well as the activity of subchondral trabecular bone were determined in osteoarthritic knees with varus or valgus deformity.

Bone specimens were taken from the lateral condyle, the medial condyle, and centrally from the intercondylar area of seven varus and four valgus knees. The percentage volume of trabecular bone was determined by histomorphometry. On an additional nine knees, five with varus and four with valgus deformity, as well as ten knees from a normal autopsy material, photon absorptiometric determination of the mineral content of the same areas was performed. On average, the loaded condyle had twice the percentage volume of trabecular bone, and accordingly twice the mineral content, of the unloaded condyle. It was remarkable that the mineral content of the latter was of the same order as the condyles of the normal material.     

The follow up study of intra-arterial infusion chemotherapy with local vein blocking as a surgical neo-adjuvant treatment for locally advanced breast cancer

Preoperative intra-arterial infusion neo-adjuvant chemotherapy, in combination with local vein blocking, was administered to thirtyone patients with locally advanced stage III breast cancer. The anti-cancer drugs and dosages used were 500 mg of 5-Fluorouracil (5FU), which was infused daily for 7–14 days, and 20 mg of Adriamycin (ADM), which was administered as a bolus dose twice into the subclavian and internal mammary arteries. The response rate of this method on the primary tumor was 48.4 per cent, and, histologically it was found to be as high as 90.3 per cent. The response rate of the clinical effects on the regional lymph nodes was 50.0 per cent, however, histologically, it was found to be lower than that of the primary tumor. In the long-term follow up study the 5-year survival rate was 72.2 per cent. Thus, this method seems to be effective as a combined modality in cases of locally advanced stage III breast cancer.     

An experimental study on the effect of vagotomy on stress ulcer and the influence of bile reflux

I investigated the significance of bile reflux, when vagotomy was done to stress ulcer, by means of measuring gastric ulcer index, gastric pH and serum gastrin levels of rats. And in order to examine influence of alkali factor of bile, I measured gastric acid output and serum gastrin levels after alkali or acid solution was infused into the untreated rat's stomach. Results were summarized as follows: 1. When I infused alkali solution into the untreated stomach, there was no significant change in serum gastrin level but gastric acid output was significantly accelerated, as compared with infusing acid solution. On the other hand, when I infused alkali solution into the vagotomized stomach, serum gastrin level increased significantly and gastric acid output was significantly accelerated, as compared with infusing acid solution, so, acceleration of acid output observed when I infused alkali solution into the untreated stomach was not suppressed in spite of vagotomy operation. 2. In the vagotomized group, gastric acid output was significantly suppressed and serum gastrin level increased significantly as compared with the untreated group. But, when bile was led into the stomach in the vagotomized group, acid output was accelerated and serum gastrin level also showed a tendency to increase as compared with the group of vagotomy alone. 3. When mild stress was inflicted, stress ulcer formation was significantly prevented in the bile reflux group (vagotomy + pyloroplasty) as well as in the non-bile reflux group (vagotomy + pyloric ligation + gastroileostomy) if vagotomy was done. But, when severe stress was inflicted, stress ulcer formation was significantly prevented in the non-bile reflux group, but not prevented in the bile reflux group. As mentioned above, it was proved that stress ulcer formation was not prevented under a severe stress, if bile reflux existed, even though vagotomy was done. So, if we do vagotomy operation, we need to choose the operation method not to induce bile reflux as much as possible.     

Perforation of gastroduodenal ulcer. Risk factors and choice of procedure

The well known surgical risk factors, such as age, time of perforation as well as size and localisation of necrosis, were confirmed in retrospective and prospective analyses of perforation of gastroduodenal ulcers (n = 504). Incidence was unchanged in the two different time-related groups, with 3.2 or 3.7 cases in one month. There was a clear-cut need for excision of the ulcer or necrotic margin. Perforation was attributable to malignant tumours in 2.1 per cent of the cases. The authors have definitely adopted the concept of earlier primary surgery (23 per cent of all cases, as compared to 10.4 per cent before). The risk associated to it was found to be lower than linked to palliative operations. The lethality rate was as low as 7.6 per cent, once due consideration was given to the above risk factors for adequate choice of surgical methods. Particular attention was given to ulcer localisation and clinical course. Hence, lethality has clearly dropped below earlier data.     

The Subchondral Bone of the Proximal Tibial Epiphysis in Osteoarthritis of the Knee

The trabecular bone of the proximal end of the tibia was assessed as an endoprosthesis-bearing structure. The mass and mineral content as well as the activity of subchondral trabecular bone were determined in osteoarthritic knees with varus or valgus deformity.

Bone specimens were taken from the lateral condyle, the medial condyle, and centrally from the intercondylar area of seven varus and four valgus knees. The percentage volume of trabecular bone was determined by histomorphometry. On an additional nine knees, five with varus and four with valgus deformity, as well as ten knees from a normal autopsy material, photon absorptiometric determination of the mineral content of the same areas was performed. On average, the loaded condyle had twice the percentage volume of trabecular bone, and accordingly twice the mineral content, of the unloaded condyle. It was remarkable that the mineral content of the latter was of the same order as the condyles of the normal material.