含银异种脱细胞真皮基质的实验研究

目的了解含银异种脱细胞真皮基质(Xeno-ADM)的多项生物学性状,观察其移植效果。方法制备单纯xeno-ADM,再用2 g／L硝酸银浸泡,制成含银xeno-ADM。检测两种xeno-ADM对笔者单位烧伤患者创面常见菌的抑菌效果,并进行组织学观察;测量含银xeno-ADM的Ag+含量。在27只家兔背部制作全层皮肤缺损创面,分为A、B、C组,每组9只。A组移植自体刃厚皮,B组移植单纯xeno-ADM+自体刃厚皮,C组移植含银xeno-ADM+自体刃厚皮。术后2、4、6周取移植部位皮肤标本作形态学观察,并计算创面收缩率;术后2周计算移植皮片(未)成活率,并检测各组家兔淋巴细胞增殖活性。结果(1)含银xeno-ADM对创面常见菌的抑菌效果明显优于单纯xeno-ADM (P<0．05)。两种xeno-ADM中的表皮已完全除去,胶原纤维粗细均匀、排列规则、无明显变性,真皮中无细胞及细胞碎片。含银xeno-ADM的Ag+含量为(2．7±0．7)mg／g。(2)术后6周,A组家兔移植皮片呈暗红色,挛缩明显,易破溃,胶原纤维排列紊乱;B、C组移植皮片颜色接近周围正常皮肤,光滑无瘢痕,质地良好,胶原纤维排列有序,表皮-真皮连接结构和基底细胞桥粒、半桥粒结构以及基底膜重建明显。术后2、4、6周,A组家兔创面收缩率均明显高于B、C组(P<0．05);B、C两组创面收缩率相似(P>0．05)。术后2周,C组皮片完全成活率为91．7％,显著高于A组(77．8％)及B组(80．6％);3组家兔淋巴细胞增殖活性相似(P>0．05)。结论含银xeno-ADM脱除了基质中有免疫原性的细胞成分,保留了组织的基本结构和完整的胶原纤维支架,且具有较好的局部抗菌效果,不失为一种良好的真皮替代物。     

异种(猪)脱细胞真皮基质与自体皮复合移植的临床应用

目的：观察基底良好的深度创面用异种（猪）脱细胞真皮基质与自体皮复合移植的临床疗效。方法：2000年1月-2002年2月用异种（猪）真皮基质与自体薄片复合移植（二步法）修复基底良好的深度创面52例。结果：37例（71.1%）植皮完全成活，15例（28.9%）植皮95%以上成活，无一例补植皮。随访3-6个月复合移植皮肤颜色、质地基本接近于正常皮肤，瘢痕增生不明显。结论：异种（猪）真皮基质可推广应用于修复基底良好的深度创面的复合移植。     

异种(猪)脱细胞真皮基质微粒与自体脂肪颗粒移植的对比研究

目的:探索一种更为理想的取材广泛的注射充填材料。方法:采取分别将异种(猪)脱细胞真皮基质微粒(xenogenicacellulardermalmatrix简称ADM微粒)和自体脂肪颗粒注入乳猪耳背皮下形成皮丘,定时对皮丘进行测量、并做必要的组织切片检查。结果:移植后24周ADM微粒吸收率约为23.86%,自体脂肪颗粒吸收率为43.77%,经方差分析P<0.01;ADM微粒注射后从外周向中央细胞化及血管化,有极少量的中性粒细胞和单核细胞。自体脂肪颗粒移植后可见到生存下来的脂肪细胞,其间有淋巴细胞和单核细胞浸润,并有异物巨细胞反应。结论:ADM微粒是一种取材广泛、安全性好、吸收率低、使用方便的理想注射充填材料。     

Control of hypertrophic scar from inception by using xenogenic (porcine) acellular dermal matrix (ADM) to cover deep second degree burn

OBJECTIVE: We have spent 7 years to investigate the method of applying porcine acellular dermal matrix (ADM) on deep partial thickness burn wound until the wound heals without dressing change. Known as "Feng's pig skin method" by our hospital, the method appears to encourage rapid re-epithilization with minimum scarring. METHOD: The deep partial thickness burn wound was rinsed cleanly under anesthesia when the patient admitted. ADM was applied on the wound after the detached epidermis was thoroughly removed, wrapped and fixed by sterile gauze and bandages. The dressing was removed within two weeks and the wound completely healed. The outcome of the treatment was analyzed by using the modified Vancouver Burn Scar Assessment Scale. RESULT: All the wounds healed with one dressing within 2 weeks, and the time of wound re-epithelialization shortened to 7-12 days. Scar hyperplasia did not occur, or it was greatly ameliorated compared with traditional treatment after a followed-up period of 3 months to 2 years. The Scar Index was significant lower than that of the traditional exposure method. CONCLUSION: Using ADM to cover deep second degree burn can preserve maximally residual dermal tissue and epithelium, help accelerate the regeneration of epithelial and stem cells, thus shorten the healing time, remodel the skin structure, and consequently has the effect of controlling hypertrophic scar at inception.     

绞窄性肠梗阻与全身炎性反应综合征相关性分析

目的探讨绞窄性肠梗阻与全身炎性反应综合征(SIRS)的关系。方法回顾性分析42例肠梗阻手术病人的临床资料。结果SIRS诊断绞窄性肠梗阻的敏感性为81.8%;特异性为85%。SIRS判断肠坏死的敏感性为95.2%;特异性为76.2%。绞窄性肠梗阻患者的白细胞明显高于单纯肠梗阻[(12.3±3.6)×109/L对(8.5±3.6)×109/L,P=0.242],心率也明显升高[(106.1±21.2)次/分对(82.3±15.1)次/分,P=0.0137]。肠坏死者的白细胞明显高于无肠坏死者[(13.4±2.9)×109/L对(9.0±2.0)×109/L,P=0.0162]。SIRS阳性者坏死肠段明显长于SIRS阴性者[(86.24±22.3)cm对5.8cm,P=0.0049]。结论SIRS能够提示肠梗阻出现肠绞窄。     

Preparation of laser micropore porcine acellular dermal matrix for skin graft: an experimental study

In our previous study, we used composite grafts consisting of meshed porcine acellular dermal matrix (PADM) and thin split-thickness autologous epidermis to cover full thickness burn wounds in clinical practice. However, a certain degree of contraction might occur because the distribution of dermal matrix was not uniform in burn wound. In this study, we prepare a composite skin graft consisting of PADM with the aid of laser to improve the quality of healing of burn wound. METHODS: PADM was prepared by the trypsin/Triton X-100 method. Micropores were produced on the PADM with a laser punch. The distance between micropores varied from 0.8, 1.0, 1.2 to 1.5mm. Full thickness defect wounds were created on the back of 144 SD rats. The rats were randomly divided into six groups: micropore groups I-IV in which the wound were grafted with PADM with micropores, in four different distances, respectively and split-thickness autograft; mesh group rats received meshed PADM graft and split-thickness autograft; control group received simple split-thickness autografting. The status of wound healing was histologically observed at regular time points after surgery. The wound healing rate and contraction rate were calculated. RESULTS: The wound healing rate in micropore groups I and II was not statistically different from that in control group, but was significantly higher than that in mesh group 6 weeks after grafting. The wound healing rate in micropore groups III and IV was lower than that in mesh and control groups 4 and 6 weeks after grafting. The wound contraction rate in micropore groups I and II was remarkably lower than that in control group 4 and 6 weeks after surgery and it was significantly much lower than that in mesh group 6 weeks after surgery. Histological examination revealed good epithelization, regularly arranged collagenous fibers and integral structure of basement membrane. CONCLUSION: Laser micropore PADM (0.8 or 1.0mm in distance) grafting in combination with split-thickness autografting can improve wound healing. The PADM with laser micropores in 1.0mm distance is the better choice.     

异种(猪)脱细胞真皮基质移植后炎症免疫反应的实验研究

目的：研究异种（猪）脱细腻真皮基质与薄自体皮复合移植后炎症反应的变化规律及对Th1/Th2细胞因子的影响。方法：将异种（猪）脱细胞真皮基质与薄自体皮复合移植于大鼠，分别于移植后1，2，3，4，8，12，16周取标本，采用组织学观察及RT-PCR技术动态检测异种（猪）脱细胞真皮基质复合移植后炎症反应及Th1/Th2细胞因子的变化，同时以单纯薄自体皮移植作对照。结果：异种（猪）脱细胞真皮与薄自体皮复合移植后早期存在炎症反应，并随着创面愈合的进行而逐渐消失，Th2细胞因子的表达明显高于对照组。结论：异种（猪）脱细胞真皮基质仍具有一定的免疫原性，Th2细胞因子的高表达说明机体对异种（猪）脱细胞真皮基质的免疫反应主要以体液免疫为主，也可能是异种（猪）脱细胞真皮基质移植后未被排斥的原因之一。     

异体颗粒状脱细胞真皮基质与自体刃厚皮复合移植修复大鼠皮肤缺损创面效果观察

目的 观察异体颗粒状脱细胞真皮基质(PADM)与自体刃厚皮复合移植修复大鼠皮肤缺损创面的效果.方法 采用随机数字表法将12只SD大鼠分为实验组和对照组,每组6只.于2组大鼠背部制作全层皮肤缺损创面,实验组创面复合移植SD大鼠异体PADM(扩张比10:5)及厚度0.20 mm的自体刃厚皮,对照组创面仅移植厚度0.20 mm自体刃厚皮.术后2周起打开敷料观察大鼠创面愈合情况.术后2、3、4、6、8、12、20周计算2组创面移植皮片成活率、收缩率(或扩张率).术后20周取2组创周正常皮肤及创面皮肤标本,采用HE染色法观察胶原纤维束结构,测量胶原纤维束直径和间隙率;用天狼星红染色法观察Ⅰ、Ⅲ型胶原分布情况,测量Ⅰ、Ⅲ型胶原含量及其比值.对实验数据行独立样本t检验、Levene检验、t'检验.结果 (1)术后2周,实验组大鼠创面移植皮片成活率[(76.1±13.1)%]低于对照组[(94.5±1.3)%,t'=3.440,P=0.018].术后3周,实验组创面移植皮片收缩率[(34±8)%]明显大于对照组[(16±12)%,t=-3.211,P=0.009];术后8周,2组移植皮片扩张率接近一致.(2)HE染色和天狼星红染色显示,与大鼠创周正常皮肤比较,对照组移植皮片胶原纤维束呈均质化改变,胶原纤维纤细,排列紊乱;实验组移植皮片胶原纤维束结构、排列更接近创周正常皮肤,可见未完全降解的PADM.与对照组创面皮肤胶原纤维束直径[(7.3±1.4)μm]、间隙率[(17±4)%]、Ⅰ型胶原含量[(68.1±8.4)%]、Ⅲ型胶原含量[(32.0±8.4)%]以及Ⅰ、Ⅲ型胶原比例(2.3±1.0)比较,实验组胶原纤维束更粗[(9.6±0.8)μm,t=-3.562,P=0.005],间隙率更大[(24±5)%,t=-2.760,P=0.020],Ⅰ型胶原含量更高[(80.2±5.4)%,t=-2.981,P=0.014],Ⅲ型胶原含量更低[(19.8±5.4)%,t=2.981,P=0.014],Ⅰ、Ⅲ型胶原比例更高(4.3±1.2,t=-3.204,P=0.009).实验组创面皮肤上述胶原相关指标更接近于创周正常皮肤水平.结论 异体PADM在体内作为真皮再生模板,有助于改善自体刃厚皮所修复的大鼠皮肤缺损创面中真皮胶原纤维束的结构,提高再生真皮组织的成熟度.

Abstract：

Objective To evaluate the effects of mixed grafting of allogeneic PADM and autologous STS on wound healing of full-thickness defect in rats. Methods Full-thickness defects with size of 6 cm×4 cm were produced on the back of 12 SD rats, and they were divided into E group(n =6) and C group ( n = 6) according to the random number table. The wounds in E group were grafted with a mix of allogeneic PADM (expansion rate 10: 5) and autologous STS with thickness of 0.2 mm, while those in C group were grafted with autologous STS in the same thickness. The wound healing rate, survival rate, contraction rate,and expansion rate of transplanted skin were observed at post operation week (POW) 2, 3, 4, 6, 8, 12,20. Tissue samples form wounds and surrounding normal skin were harvested at POW 20 for histopathological observation as follows. The structure of collagen fiber bundle was observed by HE staining, the diameter and gap rate of collagen fiber bundle were also measured. The distribution of type Ⅰ and Ⅲ collagen was observed by sirsus red staining, and the contents of type Ⅰ , Ⅲ collagen and their ratio were also examined.Data were processed with independent samples t test, Levene test, and t' test. Results Survial rate of transplanted skin in E group at POW 2 [(76. 1 ± 13. 1)%] was obviously lower than that in C group [(94.5 ± 1.3)%, t' =3.440, P =0.018]. Contraction rate of transplanted skin in E, C groups at POW 3 showed significant difference [(34±8)%vs. (16 ±12)%, t = -3.211, P =0.009]. Compared with those in peri-wound normal skin, collagen fiber bundles in C group showed signs of homogenization, and collagen fibers were thin with irregular arrangement. Collagen fiber structure and arrangement of composite skin in E group were similar to those surrounding normal skin with incomplete degradation of PADM. Diameter of collagen fiber bundle [( 9.6 ± 0.8) μm] , gap rate between collagen bundle [( 24±5) %] , content of type Ⅰ collagen [( 80.2 ± 5.4) %] and the ratio of typeⅠto type Ⅲ collagen(4.3 ± 1.2) in E group were all increased as compared with those inC group [(7.3±1.4) μm (t = -3.562, P =0.005), (17±4)%( t =-2.760, P =0.020), (68.1 ±8.4)%(t = -2.981, P =0.014), 2.3±1.0(t = -3.204, P =0. 009)], while content of type Ⅲ collagen [( 19.8 ± 5.4) %] in E group was lower than that in C group [(32.0 ±8.4)% , t = 2. 981, P = 0. 014]. Above-mentioned indexes of collagen in wound of E group were similar to those of normal skin surrounding the wound. Conclusions Allogeneic PADM used as dermal regeneration template is beneficial in improving collagen fiber bundle structure in dermis layer of rats with fullthickness skin wounds when repaired with autologous STS, and it accelerates maturation of regenerative dermal tissue.     

The effect of acellular dermal matrix on drain secretions after immediate prosthetic breast reconstruction

Abstract

In recent years the acellular dermal matrix (ADM) has gained popularity in prosthetic breast reconstruction. These procedures involve placement of a closed suction drain in the reconstructed breast. Although it is now widely accepted that ADM has an overall positive effect on the outcome of breast reconstruction, data regarding its effect on postoperative drain secretions is lacking. This study was designed to quantitatively evaluate the influence of ADM on postoperative drain secretions in the setting of immediate prosthetic breast reconstruction (IPBR). This is a prospective, comparative controlled study. Two groups of 16 patients each underwent skin sparing mastectomies (SSM) and IPBR with or without ADM. Closed suction drains were left in all the reconstructed breasts and daily secretion volumes were recorded and compared. Postoperative complications were also noted. Patients in the ADM group showed higher daily and overall secretion volumes compared with patients in the control group (p = 0.014) and the time for removal of the drains was higher by an average of 5 days (13 compared with 8 days, respectively; p = 0.004). There was no correlation between ADM and infection. This study provides the first objective evidence that ADM contributes to elevated and prolonged drain secretions when used for IPBR. This might affect possible prosthesis-related complications (e.g., rotation and malposition, capsular contraction, seroma formation, and infection). This study also noted erythema of the post-mastectomy skin flaps in selected patients, which may be attributable to a local inflammatory reaction to the ADM rather than infection.     

生态免疫肠内营养保护肠屏障功能的研究

目的探讨生态免疫肠内营养对全身炎症反应综合征(SIRS)大鼠肠屏障功能的保护作用。方法通过尾静脉注射脂多糖(LPS)建立SIRS大鼠模型,60只雄性SD大鼠随机分为4组(标准营养组、免疫增强组、生态营养组、生态免疫组),分别给予不同构成的肠内营养剂7d,观察各组大鼠血浆D-乳酸、二胺氧化酶(DAO)以及尿乳果糖/甘露醇(L/M)比值的动态变化。结果治疗7d后,生态营养组大鼠血浆D-乳酸水平明显低于标准营养组和免疫增强组(P<0.05),但这3组均高于生态免疫组3倍或以上(P<0.01)。血浆DAO水平标准营养组显著高于其他3组(P<0.01)。注射LPS后第1天起,各组大鼠尿液L/M比值均有大幅升高,至第6天回落到注射前水平。第1天时生态免疫组尿L/M比值低于其他3组(P<0.05),第3天时标准营养组尿L/M比值显著高于另外3组(P<0.01),至第6天各组已无明显差异。结论联合应用免疫增强营养素和生态制剂的生态免疫肠内营养能更有效地保护肠屏障功能。     

自体微粒皮与异体脱细胞微粒真皮混合移植对大鼠创面纤维连接蛋白和层粘连蛋白表达的影响

目的 探讨自体微粒皮与异体脱细胞微粒真皮混合移植对创面愈合的影响,并对有关机制做进一步研究.方法 Wistar大鼠作为供体,SD大鼠为受体,在SD大鼠背部建立全层皮肤损伤模型.90只SD大鼠分为5组,每组18只,第1组为自体微粒皮组;第2组为异体脱细胞微粒真皮移植组;第3、4、5组为混合移植组.混合移植组中自异体微粒皮的面积比例分别为:1∶1、1∶0.5、1∶0.25.术后第2、3、4周分别测量每组创面的愈合率,采集创面标本,做HE染色,检测纤维连接蛋白(FN)和层粘连蛋白(LN)、进行组间比较.结果 混合移植组与自体微粒皮移植组比较,混合移植组创面愈合率及FN、LN均高于自体微粒皮组,其中1∶0.25混合移植组最高,差异有统计学意义(P＜0.05或P＜0.01).结论 混合移植创面愈合率高于自体微粒皮移植,且自体微粒皮与异体脱细胞微粒真皮混合移植的面积比例按1∶0.25效果最佳,这可能与创面纤维连接蛋白和层粘连蛋白升高有关.     

Development and validation of a novel nomogram for predicting systemic inflammatory response syndrome’s occurrence in patients undertaking flexible ureteroscopy

BackgroundThe occurrence of systemic inflammatory response syndrome (SIRS) is an early alert for sepsis after flexible ureteroscopy (fURS). Once sepsis occurs, it often leads to severe or fatal consequences. We aimed to identify SIRS patients preoperatively by developing and validating a feasible prognostic nomogram model based on retrospective cohort analysis.MethodsA total of 311 patients who underwent fURS in Dongguan Kanghua Hospital (Dongguan, China) between 2016 and 2020 were included and randomly divided into a primary cohort (n=219) and validation cohort (n=92). Single factor regression analysis was used to identify the primary cohort’s meaningful characters between SIRS and non-SIRS groups. Factors of the primary cohort were then identified by least absolute shrinkage and selection operator (LASSO) regression analysis, and a nomogram was built to execute the subsequent analysis using these factors. Finally, we analyzed and drew the calibration curve, receiver operating characteristic (ROC) curve, and decision curve analysis (DCA) curve to validate the prognostic value of the nomogram in calibration and discrimination.ResultsReview of the single regression analysis of characters in the primary cohort showed gender, stone burden, diabetes, neutrophil (N), lymphocyte (L), neutrophil-to-lymphocyte ratio (NLR), platelet-to-lymphocyte ratio (PLR), lymphocyte-to-monocytes ratio (LMR), urine-WBC, nitrite (Nit), urine culture, and surgery time as significant factors between the SIRS and non-SIRS groups (P<0.05). The LASSO regression analysis suggested NLR, PLR, and urine culture were substantial factors in predicting SIRS postoperatively, lambda.min and lambda.1se (standard error, SE) were 0.01491 and 0.0796. A nomogram built with the three factors showed good calibration and discrimination, with the Brier values 0.064 and 0.034 and the area under curve (AUC) values 0.897 (95% CI: 0.837–0.957) and 0.976 (95% CI: 0.947–1.000) in the primary and validation cohort, respectively. DCA demonstrated the nomogram was clinically useful, and the predict probability of SIRS’s occurrence was very close to the actual rate as the risk threshold increased by higher than 60% in clinical impact curve analysis.ConclusionsNLR, PLR, and urine culture were significantly related to the occurrence of SIRS’s after fURS. The nomogram with these three factors showed excellent calibration, discrimination, and clinical usefulness.     

围术期容量治疗策略对心肺转流后急性炎症反应的影响

目的对比羟乙基淀粉130／0．4（万汶）与琥珀酰明胶（佳乐施）对心肺转流（CPB）后急性炎症反应的影响。方法23例择期CPB下实施冠状动脉搭桥手术（CABG）的患者随机分为两组：Ⅰ组患者的CPB预充液及术中、术后48h容量治疗选用羟乙基淀粉;Ⅱ组选用琥珀酰明胶。观察两组患者的容量治疗策略和急性炎症反应的发生情况,并分别于麻醉诱导后、停机时、术后2、24、48h采动脉血检测C反应蛋白（CRP）、白细胞介素-6（IL-6）、P选择素（P-selectin）和细胞间黏附因子（ICAM-1）含量。结果两组患者的急性炎症反应发生情况相似;术后24、48h两组CRP显著升高（P〈0．05）,但术后48hⅠ组比Ⅱ组升高更为显著（P〈0．05）;术后24、48h两组ICAM-1均较麻醉诱导后升高（P〈0．05）,而术后48hⅡ组比Ⅰ组升高更为显著（P〈0．05）。术后48hⅠ组P-Selectin、IL-6显著升高（P〈0．05）,组间差异无统计学意义。结论围术期应用羟乙基淀粉有利于激活急性期反应蛋白,并减轻内皮损伤维持内皮稳定性,从而减轻CABG后急性炎症反应。     

精子发生障碍患者精蛋白基因5′端及3′端核基质结合区的研究

运用PCR、PCR－SSCP、PCR产物直接测序等方法对61例少精子症及无精子症患者的精蛋白基因5′端及3′端核基质结合区（Matrix attachment region,MAR）进行了突变筛查。结果发现2例少精子患者在5′端核基因结合区存在PCR－SSCP带型变异,经测序分析发现其碱基序列存在一杂合性改变。