慢性光化性皮炎的病因诊断及治疗

目的;探讨慢性光化性皮炎（ＣＡＤ）的病因,诊断及治疗,方法：查阅文献,对引起ＣＡＤ的光敏物、发病机理及治疗进行分析,结果：约１１％的ＣＡＤ患者有光敏物接触史;６２％光敏性隐匿;２７％有慢性皮炎病史;结论：ＣＡＤ发病与光变态反应有关,避光治疗有效。     

钻孔减压结合自体松质骨及同种异体骨植骨治疗股骨头坏死的短期疗效观察

目的 对ARCO Ⅱa期至Ⅲa期股骨头坏死（osteonecrosis of the femoral head, ONFH）且接受大通道钻孔减压、病灶清除,并经减压通道行股骨头内自体松质骨、同种异体颗粒骨打压植骨、同种异体腓骨条植骨治疗的患者进行短期临床疗效分析。方法 2018年1月至2020年12月,四川省骨科医院纳入47例（59髋）股骨头坏死患者。其中,男39例,女8例;年龄29 ～ 51岁,平均37.2岁;ARCO Ⅱa期16髋,ARCO Ⅱb期21髋,ARCO Ⅱc期13髋,ARCO Ⅲa期9髋;酒精性ONFH 46髋（酒精组）,激素性ONFH 13髋（激素组）。术后门诊随访,复查X线片（双髋正位及蛙式位片）,观察股骨头形态及骨整合情况,记录患髋Harris评分。结果 本组47例（59髋）随访24 ～ 36个月,平均随访30个月。术后所有患者疼痛明显缓解,部分患者疼痛消失（包括Ⅱa期14髋,Ⅱb期13髋,Ⅱc期4髋）。6例（9髋）患者病情进展股骨头发生塌陷,其中Ⅱb期1髋,Ⅱc期3髋,Ⅲa期5髋。按病因分,酒精性坏死塌陷6髋,塌陷率13%;激素性坏死塌陷3髋,塌陷率23.1%。酒精组和激素组末次随访时Harris评分与术前比较,差异均有统计学意义（P＜0.05）;酒精组术后Harris评分（82.7±5.6）分显著高于激素组（78.3±8.3）分,差异具有统计学意义（P＜0.05）。不同ARCO分期患者术后髋关节功能均得到一定程度改善,其中Ⅱa及Ⅱb期优良率78.4%,Ⅱc、Ⅲa期优良率50%（P＜0.05）。结论 针对ARCO Ⅱa期至Ⅲa期股骨头坏死的患者,采用大直径通道减压、清除死骨,同时植入自体松质骨和同种异体颗粒骨混合物,最后配合同种异体腓骨条加强支撑,短期可有效防止坏死股骨头的塌陷并减轻疼痛、改善患功能。     

4R问题分析法结合反向教学在小儿手术室护理教学中的应用

目的 探讨4R问题分析法结合反向教学在小儿手术室护理教学中的应用。方法 本研究将2020年8月至2022年8月分配到湖南省儿童医院手术室实习的护生480人作为研究对象,按进科先后顺序分为对照组和观察组,每组240人。对照组采用传统教学,观察组采用4R问题分析法结合反向教学。实习结束后比较两组实习护生的综合能力考核成绩、护理教学认可度及教学满意度。采用SPSS 22.0进行t检验和卡方检验。结果 观察组实习护生理论知识、基本护理技能、手术室专科护理技能及综合护理技能考核成绩得分均明显高于对照组,差异有统计学意义（P<0.05）。观察组实习护生护理教学认可度明显优于对照组,差异有统计学意义（P<0.001）。两组实习护生教学满意度比较差异有统计学意义（χ²=118.35,P<0.001）。结论 4R问题分析法结合反向教学可有效提高小儿手术室实习护生的护理教学质量和教学满意度,使实习护生更好地掌握理论和实践技能。     

重组汉坦病毒核壳蛋白的制备及其单克隆抗体的研制

将ＰＣＲ扩增的ＨＴＶ７６－１１８ＲＮＡ小片段的ｃＤＮＡ克隆插入到载体ｐＤＳ５６／ＲＢＳⅡ－（Ｏ）－６Ｈｉｓ中,通过ＩＰＴＧ诱导表达以及镍螯合层析纯化重组ＮＰ,经过ＳＤＳ－ＰＡＧＥ、免疫转印和ＥＬＩＳＡ方法分析其蛋白特性,证实该重组ＮＰ与毒粒ＮＰ相同,均为４９．６ｋｕ,能与肾综合征出血热（ＨＦＲＳ）患者血清产生特异性反应。以此重组ＮＰ作为抗原制备抗ＮＰ单克隆抗体,获得两株持续稳定分泌单克隆抗体的杂交瘤细胞株。用抗ＮＰＭｃＡｂ建立检测ＩｇＭ抗体的ＥＬＩＳＡ捕获法。证实纯化重组ＮＰ作为抗原,避免了组织培养病毒的感染性和非特异性以及难以标准化的缺点。所建立的ＥＬＩＳＡ捕获法检测ＨＦＲＳ患者血清中ＩｇＭ抗体的方法特异性强、敏感性高,可作为ＨＦＲＳ的早期诊断试剂     

用PCR体外定点突变技术诱导霍乱毒素A亚基突变体的构建

目的： 有效去除霍乱毒素Ａ 亚基（ ＣＴ Ａ） 的毒性作用而保存其佐剂性能。方法： 采用ＰＣＲ 体外定点突变技术（ＰＣＲ ＳＤＭ） ,设计两对引物,引入一个突变位点,通过重叠延伸法两次ＰＣＲ 扩增,使ＣＴ Ａ 编码基因第６３ 位密码子由ＣＴＣ 突变为ＴＴＴ,亦将扩增片断克隆入ＰＵＣ１９ 载体。结果： ＤＮＡ 测序结果表明在预期位点已发生突变,用ＰＣＲ 诱导成功ＣＴ Ａ 突变体。结论： ＰＣＲ 诱导突变准确、简便,为深入研究ＣＴ 免疫佐剂的作用打下了基础     

用LDH释放法判断细胞病变程度确定干扰素生物活性

目的：应用ＬＤＨ释放法测定干扰素生物活性。方法：通过检测ＷＩＳＨ细胞裂解后释放出的ＬＤＨ活性判断细胞病变的程度,确定干扰素效价。对ＬＤＨ释放法进行改进,与结晶紫染色法和ＭＴＴ比色法进行灵敏度、重复性和相关性比较。结果和结论：该法减少了实验步骤和时间,成本低,敏感性高,重复性好,与其他方法所测结果无显著差异,简便实用,优于结晶紫染色法和ＭＴＴ比色法。     

Expression of the extracellular matrix proteins collagen I, collagen III and fibronectin and matrix metalloproteinase-1 and -13 in the skin of patients with inguinal hernia

Although abnormal collagen metabolism has been ascribed an important role in the high recurrence rates after surgical hernia repair, knowledge on tissue sampled in the region affected by inguinal hernias is poor. In the present study, we determined collagen type I and type III in the skin of adult patients with indirect and direct inguinal hernias by both immunohistochemistry and Western blot analysis. In addition, we quantified the immunohistochemical expression of fibronectin and matrix metalloproteinase (MMP)-1 and -13. The results indicated that the ratio of collagen type I/III was significantly decreased in the skin of patients with either indirect (n = 9) or direct hernia (n = 7), with a concomitant increase in collagen type III (p < 0.001 vs. controls, n = 7, without affection of the inguinal region). There was no significant difference between patients with indirect and direct hernia (p > 0.05). MMP-13 was not expressed in any of the skin samples investigated, whereas MMP-1 was found in the epidermis. Fibronectin was predominantly detected at the epidermal-dermal junction. MMP-1, MMP-13 and fibronectin levels were significantly different between patients and controls (p > 0. 05). We conclude that in contrast to the unchanged expression of fibronectin and MMP-1 and MMP-13, the decreased ratios of collagen tpye I/III with the basically increased amount of collagen type III could be of significant importance for the pathophysiology of hernias. The specific ratio collagen I/III probably reflects the altered structural integrity and mechanical stability of the connective tissue in both indirect and direct hernias. Moreover, our findings stress that hernias should be regarded as the manifestation of a systemic disease in the inguinal region with a genetic background, explaining the high recurrence rates after repeated suture repair, as well as the usefulness of surgical meshes in this clinical setting.     

Synthesis of type I and III collagen, expression of fibronectin and matrix metalloproteinases-1 and -13 in hernial sac of patients with inguinal hernia

Since many years the importance of a weakness of the soft tissue for the development of hernias is discussed controversially. The tensile strength of the tissue is supposed to depend largely on the varying proportion of type I collagen with its high tensile strength and the immature type III collagen. Their relation is regulated by several collagenases, mainly matrix metalloproteinases-1 and -13 (MMP-1 and MMP-13), whereas fibronectin plays a key role for the adherence of cells within the extracellular matrix. The aim of this study was to investigate whether an alteration in type I and type III collagen synthesis, amounts of MMP-1 and MMP-13 and the expression of fibronectin were associated with the development of inguinal hernia. We analysed the hernial sac of patients with indirect (n = 9) and direct (n = 7) inguinal hernias and peritoneum in controls (n = 7) by immunohistochemistry and Western blot analysis. The results showed that the ratio of relative amount of I/III collagen was markedly decreased in patients with either indirect or direct hernias as compared with controls (p < 0.001) with a concomitant increase in type III collagen synthesis. MMP-13 was expressed neither in the hernial sac nor in the peritoneum of the controls, but the positive reactions of MMP-1 were found in the surface of the subserosa of the hernial sac in patients with indirect or direct hernias without any difference compared to controls. Furthermore, the relative amount of fibronectin in patients with either indirect or direct hernias is not significantly different from controls (p > 0.05). In regard to the known alterations of the collagen metabolism in fascia and skin of hernia patients the changed collagen I/III ratio with its increase of type III collagen in hernial sacs support the presence of a systemic disturbance of collagen metabolism. The absence of changes of the expression of collagenases (MMP-1, MMP-13) and the constant levels of fibronectin underline the central role of collagen synthesis for the development of indirect or direct hernias.