Induction of endometrial mesenchymal stem cells into tissue-forming cells suitable for fascial repair

Pelvic organ prolapse is a major hidden burden affecting almost one in four women. It is treated by reconstructive surgery, often augmented with synthetic mesh. To overcome the growing concerns of using current synthetic meshes coupled with the high risk of reoperation, a tissue engineering strategy has been developed, adopting a novel source of mesenchymal stem cells. These cells are derived from the highly regenerative endometrial lining of the uterus (eMSCs) and will be delivered in vivo using a new gelatin-coated polyamide scaffold. In this study, gelatin properties were optimized by altering the gelatin concentration and extent of crosslinking to produce the desired gelation and degradation rate in culture. Following cell seeding of uncoated polyamide (PA) and gelatin-coated meshes (PA + G), the growth rate of eMSCs on the PA + G scaffolds was more than that on the PA alone, without compromising cell shape. eMSCs cultured on the PA + G scaffold retained their phenotype, as demonstrated by W5C5/SUSD2 (eMSC-specific marker) immunocytochemistry. Additionally, eMSCs were induced to differentiate into smooth muscle cells (SMC), as shown by immunofluorescence for smooth muscle protein 22 and smooth muscle myosin heavy chain. eMSCs also differentiated into fibroblast-like cells when treated with connective tissue growth factor with enhanced detection of Tenascin-C and collagen type I as well as new tissue formation, as seen by Masson’s trichrome. In summary, it was demonstrated that the PA + G scaffold is an appropriate platform for eMSC delivery, proliferation and differentiation into SMC and fibroblasts, with good biocompatibility and the capacity to regenerate neo-tissue.     

腰椎间盘突出症术中神经根鞘内及其周围用药疗效研究

背景　传统的骶管注射或术中使用神经阻滞药物是治疗腰椎间盘突出症（LDH）或术后早期炎性水肿引起的神经根性疼痛的有效方法。然而,用于神经根鞘内及其周围的神经阻滞药物作用时间短,疗效有限,仍有进一步研究的必要性。目的　观察LDH术中神经根鞘内及其周围用药的临床疗效。方法　根据纳入与排除标准,选取2015年3月-2018年3月于广西中医药大学第一附属医院行椎板开窗髓核摘除、神经根管扩大术治疗的LDH患者90例。采用随机数字表法将患者分为A组（n=30,给予神经根鞘内及其周围用药）、B组（n=30,给予神经根鞘内用药）、C组（n=30,不做特殊处理）。收集患者一般资料、术后卧床时间、术后住院时间、术后神经根性疼痛情况;记录术前及术后第1、3、6、10天视觉模拟评分法（VAS）评分,第3、6、10天日本骨科学会治疗评估评分法（JOA）评分;观察不良反应发生情况。结果　B组、C组术后卧床时间、术后住院时间长于A组,术后神经根性疼痛发生率高于A组（P<0.05）;C组术后卧床时间、术后住院时间长于B组,术后神经根性疼痛发生率高于B组（P<0.05）。干预方式与时间在VAS评分上存在交互作用（P<0.05）;干预方式、时间在VAS评分上主效应显著（P<0.05）。B组、C组术后第1、3、6、10天VAS评分高于A组（P<0.05）;C组术后第1、3、6、10天VAS评分高于B组（P<0.05）。A组、B组、C组术后第1、3、6、10天VAS评分均低于本组术前（P<0.05）。干预方式与时间在JOA评分上存在交互作用（P<0.05）;干预方式、时间在患者JOA评分上主效应显著（P<0.05）。B组、C组术后第6、10天JOA评分低于A组（P<0.05）;C组术后第6、10天JOA评分低于B组（P<0.05）。A组、B组、C组术后第3、6、10天JOA评分均高于本组术前（P<0.05）。3组患者均无明显不良反应。结论　LDH术中神经根鞘内及其周围用药可能通过提高药物的作用时间来延长镇痛时间,促进LDH术后早期恢复,减轻术后神经根性疼痛,减少卧床时间及住院时间,提高术后疗效。     

Gelatin-based hemostatic agents for medical and dental application at a glance: A narrative literature review

Uncontrolled bleeding is linked to higher treatment costs, risk of post-surgical infection and increased disease and death. Hemostatic agents are used to treat excessive bleeding. A good hemostatic agent controls bleeding effectively, reduces the need for blood transfusion, removes the need for systemic drugs to control bleeding, results in shorter surgery time, and reduces the cost and length of hospital stay of the patient. Gelatin-based hemostatic agents have been widely used in medical and dental procedures, owing to their biodegradability and biocompatibility, as well as availability and low cost of raw materials. In this narrative literature review, we discuss the background and different types of gelatin-based hemostatic agents in medical and dental procedures, the comparison of gelatin-based and non-gelatin-based hemostatic agents, and the usage and development of enhanced or novel gelatin-based hemostatic agents. Gelatin-based hemostatic agents are effective and important part of bleeding control, as evidenced by its wide application in medicine and dentistry. The development of novel combination gelatin-based hemostatic agents has much potential for effective control of excessive bleeding.     

肠球菌部分致病基因和表型的检测

目的 调查临床分离的肠球菌6种致病基因和2种表型的分布。方法 应用聚合酶链反应和斑点杂交技术检测临床分离的145株肠球菌的6种致病基因，用7％兔血平板和3vA，明胶平板检测B溶血和明胶溶解表型。结果 粪肠球菌和屎肠球菌6种致病基因的检出率分别为gelE 72．9％、30．6％；efaA 79．2％、36．7％；cylA 54、2％、34．7％；esp 34、4％、36．7％；agg 18．8％、0；ace28．1％、0。粪肠球菌和屎肠球菌β溶血分别为45．8％、20．4％；明胶溶解分别为35、4％、16．3％。结论 粪肠球菌6种致病基因和2种表型的检出率高于屎肠球菌；尿标本比痰标本肠球菌致病基因的检出率高；gelE、efaA和cylA在粪肠球菌中的检出率高于其他致病基因。     

改良固定底物膜法对精子顶体透明质酸酶检测的研究

目的研制一种检测人精子透明质酸酶的改良底物膜法，以提高临床对男性不育的诊断水平。方法根据精子顶体透明质酸酶能溶解卵丘基质透明质酸的生化特性，通过改良透明质酸钠-明胶底物膜法，后进行孵育及染色来显示透明质酸酶的活性。本研究检测了70例人精子透明质酸酶活性，并结合临床精液常规检查结果选择和分组标本（生育组26例，不育A组13例，不育B组31例），对生育组与各不育组之间的平均酶活性进行统计学分析。结果在普通光镜下即可观察到在阳性反应区以深紫蓝色底膜为背景的清晰明亮的消化晕环，晕环数量及直径的大小与透明质酸酶活性成正比。生育组透明质酸酶平均阳性率为70．84％，酶活性亮区平均直径为78．17μm；不育A组酶平均阳性率为60．02％，酶活性亮区平均直径为76．92μm；不育B组酶平均阳性率为29．11％，酶活性亮区平均直径为8．22μm；经t检验，生育组酶活性两项指标均高于不育B组，差异有统计学意义（P〈0．01）。结论本研究检测方法简便、可靠，并可连续观察单个精子顶体透明质酸酶活性反应，很适合作为临床评价精子功能的有效指标，并提高对男性不育的诊断水平。     

腰-硬联合麻醉时快速输注琥珀酰明胶联合麻黄碱静脉滴注预防剖宫产术中低血压

目的观察腰-硬联合麻醉时快速输注琥珀酰明胶扩容联合麻黄碱静脉滴注预防剖宫产术中低血压的效果。方法选择90例ASAⅠ~Ⅱ级在腰-硬联合麻醉下行剖宫产孕妇,采用随机数字表法随机分为A、B、C组各30例。均于腰麻前10分钟开始快速输液,30分钟内各输入液体量10 ml/kg,A组输入复方氯化钠液,B组输入琥珀酰明胶,C组输入琥珀酰明胶并于产妇平卧后静脉注射麻黄碱10 mg。输药之后,3组均以10 ml/(kg.h)的速度输注乳酸林格氏液维持。对比观察3组麻醉手术期间血压、心率的变化,3组低血压的发生率及不良反应的发生情况。结果 C组麻醉前后血压心率变化小;血流动力学比A、B组稳定(P<0.01或0.05),低血压及恶心呕吐的发生率也明显少于A、B组(P<0.05)。B组血压下降程度及低血压发生率虽小于A组,但组间比较差异无统计学意义(P>0.05)。结论腰-硬联合麻醉期间快速输注琥珀酰明胶扩容联合麻黄碱静脉滴注能有效预防剖宫产术中低血压的发生。     

时间分辨免疫荧光法用于梅毒抗体测定的探讨

目的 通过对时间分辨免疫荧光法（TRFIA）,酶联免疫吸附试验（ELISA）,明胶颗粒凝集试验（TPPA）三种方法测定梅毒特异性抗体（TP）的比较,并对TRFIA法的特异性、灵敏度、线性进行方法学评价,探讨临床常规使用TRFIA方法检测梅毒特异性抗体的可行性.方法 对2012年1月至2012年7月我院256名门诊患者标本分别用TRFIA、ELISA、TPPA三种方法测定TP,并对TRFIA检测系统的灵敏度、批内,批间精密度、线性范围进行测定.结果 TRFIA与TPPA法检测相比其灵敏度为100％,特异性为98.2％.TP阴阳性结果,经配对卡方检验P＞0.05,三种方法无统计学差别.最低检测浓度0.15NCU,相关系数分别为0.95、0.999,高、中、低三种浓度血清的批内、批间精密度CV＜10％.结论 TRFIA法测定TP灵敏度、特异性、方法性能、相关性均好,可作为临床TP常规筛查方法.     

非特异性梅毒螺旋体抗体对HIV明胶颗粒凝集试验的影响

目的 探讨梅毒血清中非特异性梅毒螺旋体抗体对艾滋病病毒(HIV)明胶颗粒凝集试验的影响.方法 采用回顾性调查的方法,用快速血浆反应素环状卡片试验(RPR)、HIV明胶颗粒凝集试验(HIV1/2-PA)、梅毒螺旋体明胶颗粒凝集试验(TPPA)以及Abbot HIV胶体硒快速试验(Determine HIV-1/2),同时检测性病门诊保存的96份梅毒血清及96份健康对照者血清.结果 96份梅毒血清中有32份HIV1/2-PA结果被干扰而无法判读(致敏颗粒和非致敏颗粒同时凝集),其他64份血清HIV1/2-PA结果为阴性.将96份梅毒血清按滴度分成11组,观察到随着RPR滴度的升高,HIV1/2-PA结果出现干扰现象的频率也升高.RPR滴度＞1:8的标本出现此现象的比例与≤1:8的标本相比,两者差异有统计学意义(χ2=74.73,P＜0.005).对9例出现HIV1/-PA结果干扰的梅毒患者进行随访,在苄星青霉素规范治疗后6个月,用HIV1/2-PA测试其血清,原结果中的干扰现象消失.比较这9例患者治疗前后RPR和TPPA滴度的变化发现,RPR的滴度均下降至≤1:8,而TPPA的滴度则有4例升高,有4例未变化.5份RPR阳性、滴度均＜1:8.TPPA阴性标本,在HIV1/2-PA试验中未出现结果被干扰现象.结论 高滴度(＞1:8)的非特异性梅毒螺旋体抗体,是HIV1/2-PA出现致敏颗粒和非致敏颗粒同时凝集这一干扰现象的主要原因.     

Preventive effects of chitosan on peritoneal adhesion in rats

AIM: To study the effects of chitosan gel and blending chiston/gelatin film on preventing peritoneal adhesion in rats. METHODS: SD rats were randomly divided into 2 groups, group A treated with chitosan gel and group B with blending chiston/gelatin film. In group A, rats were randomly subdivided into 3 subgroups as groups A1, A2 and A3, and different methods were used to induce peritoneal adhesions at the dead end of vermiform process in each group as follows: Group Al with trauma, A2 with talc powder and A3 with ligation of blood vessel. In each subgroup, rats were redivided into control group and experimental group whose treated vermiform processes were respectively coated with chitosan gel and normal saline immediately after the adhesion-induced treatments. In group B, all the rats received traumatic adhesion-induced treatments and then were randomly divided into 4 groups (groups B1, B2, B3, B4). Group Bl served as control group and were coated with normal saline in the vermiform processes immediately after the treatments, and groups B2, B3 and B4 with 100% chitosan film, chitosan film containing 10% gelatin and chiston film containing 50% gelatin, respectively. At 2 and 4 wk after the above treatments, half of the rats in each terminal group were belly opened, and the peritoneal adhesive situation was graded and histopathological changes were examined. RESULTS: (1) In group A, regarding peritoneal adhesion situation: At both 2 and 4 wk after the treatments, for groups Al and A3, the adhesive grades of experimental groups were significantly lower than those of the control group (2 wk: H = 4.305, P < 0.05 for Al, H = 6.743, P < 0.01 for A3; 4 wk: H = 4.459, P < 0.05 for A1, H = 4.493, P < 0.05 for A3). However, of group A2, there was no significant difference between the experimental and control groups (2 wk: H = 0.147, P > 0.05; 4 wk: H = 1.240, P > 0.05). Regarding pathological changes: In groups A1 and A3, the main pathological change was fibroplasia. In group A2, the main changes were massive foreign-body giant cell reaction and granuloma formation with fibroplasia of different degrees. (2) In group B, regarding degradation of film: With increase of the blended gelatin concentration, degrading speed of the film accelerated significantly. Regarding peritoneal adhesion situation: At both 2 and 4 wk after the treatments, the adhesive grades of B1 were the lowest among the four subgroups of B (2 wk: H = 29.679, P < 0.05; 4 wk: H = 18.791, P < 0.05). At 2 wk after the treatments, the grades of group B2 were significantly lower than that of groups B3 and B4 (H = 4.025, P < 0.05 for B2 vs B3; H = 4.361, P < 0.05 for B2 vs B4). At 4 wk, there were no significant differences of the grades between groups B2, B3 and B4. Regarding pathological changes: Inflammatory cell infiltration and fibroplastic proliferation were observed in the local treated serous membranes, which was the mildest in group B1. Slight foreign-body giant cell reactions were also found in groups B2, B3, and B4. CONCLUSION: (1) Chitosan gel has preventive effect on traumatic or ischemic peritoneal adhesion, but no obvious effect on foreign body-induced peritoneal adhesion. (2) Chitosan film may exacerbate the peritoneal adhesion. Blending with gelatin to chitosan film can accelerate the degradation of the film, but can simultaneously facilitate the formation of peritoneal adhesion.