封闭负压引流对人慢性创面中基质金属蛋白酶的影响

目的：研究人慢性创面在经封闭负压引流（Vacuum-assisted-closure V.A.C)治疗前后，创面肉芽组织中基质金属蛋白酶（metalloproteinase,MMp)MMP1,MMP2,MMP13，以及金属蛋白酶组织抑制剂（tissue inhibitor of melalloproteinase,TIMP)TIMP-1,TIMP-2 mRNA的变化。方法：对9例慢性创面患者给予V.A.C治疗（－120mmHg压力），分别于治疗前和治疗后1，4，7天切取创面中央肉芽组织，提取总RNA，利用RT－PCR方法测定MMP－1，MMP－2，13和TIMP－1，TIKMP－2mRNA的表达情况，并对其定量结果进行统计学分析。结果；MMP－1，13mRNA在V.A.C治疗后表达下降，以MMP－13下降趋势尤为明显。MMP－2mRNA表达呈现波动，但总体为下降趋势，在V．A．C治疗前，检测不到TIMP－1，2mRNA的表达，应用V．A．C后，出现了两者的表达，而且随时间的延长，TIMKP－1，2mRNA的表达呈现明显的上升趋势。结论：V．A．C通过抑制MMP－1，2，13mRNA的表达，促进TIMP－1，2mRNA的表达，使更多的MMP与TIMP形成复合物进而抑制胶原和明胶的降解，促进慢性创面的愈合，同时也反向证明慢性创面形成过程中，MMP－1，2，13mRNA的表达增加以及TIMP－1，2，mRNA表达下降是重要因素。     

封闭负压引流技术对创面愈合过程中原癌基因表达的影响

目的 研究封闭负压引流技术(vacuum-assisted closure，VAC)对启动创面愈合过程和减少细胞凋亡的作用。方法 用免疫组化法检测猪急性皮肤缺损创面和人慢性创面原癌基因c-myc、c-jun和Bcl-2表达变化，计算阳性表达细胞数和标记指数，观察创面愈合过程。结果 ①VAC治疗组猪急性皮肤缺损创面清洁无明显渗出，第6天即有较多新生上皮和肉芽组织，25d完全愈合。对照组创面有较多渗出和血痂，第6天出现少量新生上皮和肉芽组织，30d愈合。伤后即刻c-myc、c-jun和Bcl-2表达量少，主要位于基底细胞的胞浆或胞核，伤后及VAC治疗后表达迅速显著增加，但表达至峰值后迅速下降。在伤后或VAC治疗后的12d内实验组表达始终高于对照组。②人慢性创面VAC治疗后分泌物明显减少，较快形成健康的肉芽组织。c-jun主要表达在表皮基底细胞、真皮成纤维细胞和炎性细胞的胞浆，VAC治疗后阳性细胞数和标记指数显著减少。c-myc和Bcl-2主要表达在基底细胞的胞浆，VAC治疗后表达量显著增多，标记指数显著增大。结论 VAC能快速启动猪急性皮肤创面和人慢性创面的愈合过程，减少修复细胞凋亡，使创面愈合加速。     

封闭负压引流技术对失感觉神经支配创伤愈合中Bcl-2与NGF/NGF mRNA表达的影响

目的　研究封闭负压引流技术 (vacuum assistedclosure ,VAC)对失感觉神经支配的创面愈合过程中Bcl 2与NGF表达的影响。方法　将 80只SD大鼠随机分成四组 (每组 2 0只 ) ,即实验组(T组 ) :施加VAC的失去神经支配创面 ;对照组 1(C1) :未施加VAC的失去神经支配的创面 ;对照组2 (C2 ) :施加VAC的正常神经支配的创面 ;对照组 3(C3) :未施加VAC的正常神经支配的创面。T、C2组施加间断性VAC每日 3次 ,负压 80mmHg。伤后 1、3、6、9、12d ,四组同时取材 ,采用免疫组化和原位杂交检测各组各时间点Bcl 2与NGF/NGFmRNA表达。结果　在伤后C2组、C3组Bcl 2与NGF/NGFmRNA表达明显 ,水平逐渐升高 ,至第 9天达最高 ,然后降低 ,C2组较C3组Bcl 2和NGF/NGFmRNA维持相对高的状态 (P <0 .0 5 )。T、C1组创缘和肉芽组织中Bcl 2与NGF/NGFmRNA表达相对C2、C3组处于较低水平 (P <0 .0 5 )。结论　VAC通过增强创面组织抑制凋亡相关基因蛋白的表达 ,影响内源性NGF表达 ,具有明显的促进创面愈合的作用。     

基质金属蛋白酶和组织金属蛋白酶抑制剂在大肠癌中的表达

目的　研究基质金属蛋白酶 (MMPs)及组织金属蛋白酶抑制剂 (TIMPs)在大肠癌中的表达特点 ,与肿瘤发生发展的关系 ,以及在肿瘤治疗中的应用前景。　方法　采用RT PCR方法测定 2 8例大肠癌患者肿瘤组织和周围正常粘膜的基质金属蛋白酶 2 (MMP 2 )、膜型 1 基质金属蛋白酶 (MT1 MMP)、基质溶素 (MMP 7)、组织金属蛋白酶抑制剂 2 (TIMP 2 )、组织金属蛋白酶抑制剂 3(TIMP 3)的mRNA表达状况 ,并将其结果与临床及病理学资料进行统计学分析。　结果　 (1) 2 7例患者肿瘤组织中MMP 7mRNA表达阳性 ,MMP 2、MT1 MMP、TIMP 2和TIMP 3在肿瘤组织和正常粘膜中均有高表达 ;(2 )肿瘤组织中MMP 7mRNA的表达水平与大肠癌患者的Dukes′分期相关 (P <0 0 1) ;(3)淋巴结阳性患者的肿瘤组织TIMP 2表达水平为 (1 2 5± 0 46 )明显高于淋巴结阴性患者的 (0 75± 0 41) ,差异有显著性意义 (P <0 0 1) ;(4)大肠癌患者癌周正常粘膜TIMP 3mRNA表达随患者Duke′s分期的进展和肿瘤浸润深度的增加而降低 (P <0 0 1) ;(5 )TIMPs与MMPs之间无明显相关关系 (P >0 1)。　结论　MMP 7可望成为诊断大肠癌的敏感指标 ;人工诱导TIMP 2、TIMP 3或阻断MMP 7、MMP 2、MT1 MMP的表达可能抑制肿瘤的浸润和转移 ,成为肿瘤治疗的新途径。     

膀胱癌组织MMP-9和TIMP-1的表达及其意义

目的　研究金属蛋白酶 9(MMP 9)及其抑制因子 (TIMP 1)在膀胱癌中的表达及其临床意义。方法　应用免疫组化方法检测MMP 9及TIMP 1在 6 5例膀胱癌和 19例非肿瘤性膀胱组织中的表达。结果　MMP 9在膀胱癌细胞和间质组织中均有表达 ,而TIMP 1主要表达在膀胱癌细胞中 ,在间质组织中很少表达。TIMP 1在膀胱癌细胞中的表达低于非肿瘤性膀胱组织中移行上皮的表达 ,两者差别有统计学意义 (P <0 .0 5 ) ,但与膀胱癌的病理分级、分期无统计学相关性 (P >0 .0 5 )。MMP 9在膀胱癌细胞及间质中的表达均高于非肿瘤性膀胱组织 ,两者差别有统计学意义 (P <0 .0 5 ) ,而且与膀胱癌的病理分级、分期均呈正相关 (P <0 .0 5 )。膀胱癌细胞表达TIMP 1与间质组织表达MMP 9呈负相关 (P <0 .0 5 ) ,MMP 9及TIMP 1的表达与肿瘤的复发、多发均无相关性 (P >0 .0 5 )。结论　MMP 9和TIMP 1参与膀胱癌的浸润和转移 ,对预后判断、诊断及治疗具有一定的意义。     

生长抑素对大鼠移植性肝癌VEGF、MMP的影响

目的　观察生长抑素对大鼠移植性肝癌血管内皮生长因子 (VEGF)、基质金属蛋白酶 1(MMP 1)表达的影响 ,初步探讨生长抑素抑制大鼠移植性肝癌生长的机制。方法　制作大鼠移植性肝癌的动物模型 ,随机分为 2组 ,对照组 (n =11) ,治疗组 (n =8)。治疗组腹腔注射善宁 (10 0 μg·kg-1·d-1) ,每天分 2次注射 ,对照组注射等量生理盐水。 10d后心脏采血 0 5ml后处死大鼠 ,大鼠血液室温下 4h ,待其凝固 ,离心 (30 0 0转 /min) 10min ,分离血清放入 - 80℃冰箱保存。酶联免疫吸附试验检测血清VEGF ,原位杂交法检测瘤组织VEGF、MMP表达。结果　治疗组血清VEGF值低于对照组 ,瘤组织VEGFmRNA、MMP 1mRNA在对照组、治疗组中均有阳性表达 ,而治疗组表达水平较对照组低 (P <0 0 5 )。结论　生长抑素抑制血清VEGF ,转移相关基因VEGF、MMP 1表达降低 ,可能是生长抑素抑制大鼠移植性肝癌生长的机制之一。     

碱性成纤维细胞生长因子对大鼠烫伤创面中基质金属蛋白酶及其抑制剂的影响

目的　观察大鼠深Ⅱ度烫伤创面愈合过程中基质金属蛋白酶 (MMP) 2、MMP 7与金属蛋白酶组织抑制因子 2 (TIMP 2 )的变化以及创面外用碱性成纤维细胞生长因子 (bFGF)对其的影响。　方法　制作 30 %TBSA深Ⅱ度烫伤大鼠模型 ,分为单纯烫伤组和bFGF治疗组。于伤后 3、6h和 1、3、7、14d取创面皮肤标本 ,检测再上皮化率及胶原含量 ,并采用免疫组织化学染色法检测创面组织真皮内成纤维细胞MMP 2、MMP 7和TIMP 2的表达变化。另取 6只正常大鼠作为假烫组 ,分别检测上述各项指标。　结果　 (1)伤后 3～ 14d ,bFGF治疗组再上皮化率高于单纯烫伤组。 (2 )伤后 3h～ 3d ,bFGF治疗组和单纯烫伤组胶原含量持续下降 ,7～ 14d开始回升 ,但仍低于假烫组 (P<0 .0 5 )。 (3)伤后 1d ,单纯烫伤组MMP 2、MMP 7和TIMP 2表达增多 ,7d时达到高峰 ,持续到 14d。(4 )伤后 3～ 6h,bFGF治疗组MMP 2、MMP 7的表达与单纯烫伤组相似 ;伤后 1～ 14d ,3者的阳性表达强于单纯烫伤组。　结论　烫伤大鼠创面中细胞外基质的活动会影响皮肤的胶原沉积 ;MMP 2、MMP 7、TIMP 2的表达变化是组织修复的重要步骤 ,与bFGF加速创面愈合的过程密切相关。     

深Ⅱ度烫伤大鼠创面基质金属蛋白酶-1,2及其抑制剂表达变化机制的探讨

目的　观察深Ⅱ度烫伤创面愈合过程中基质金属蛋白酶 1,2 (MMP 1,2 )与金属蛋白酶组织抑制因子 1,2 (TIMP 1,2 )的表达规律以及其变化机制。　方法　 150只Wistar大鼠 3 0 %深Ⅱ度烫伤后随机分为 5组 :(1)正常对照组 (n =6) ;(2 )单纯烫伤组 (n =3 6) ;(3 )激酶激活剂BDM组 (n =3 6) ;(4)蛋白激酶C制剂H7组 (n =3 6) ;(5)c fos抗体组 (n =3 6)。分别于伤后 3、6h和 1、3、7、14d采取创面皮肤标本 ,检测创面组织c fosmRNA与c fos蛋白的表达 ,以及MMP 1,2 /TIMP 1,2相应变化。　结果　伤后 3～ 6h ,c fosmRNA与c fos蛋白的表达明显增多 ,随后逐渐下降。MMP 1,2 /TIMP 1,2的表达滞后于前者 ,在伤后 3～ 7d表达较高 ,MMP 2 /TIMP 2的表达增加显著。使用BDM后 ,烫伤 3～6h组织c fosmRNA与c fos蛋白的表达增加 ,MMP 1,2 /TIMP 1,2的表达也增强。使用H7后 ,c fosmRNA与c fos蛋白的表达被抑制 ,MMP 1,2 /TIMP 1,2的表达随之减弱。c fos抗体中和内源性c fos蛋白表达后 ,MMP 1/TIMP 1,2的表达受抑制 ,MMP 2的变化不明显。　结论　创面愈合过程中 ,MMP 1,2和TIMP 1,2的表达与蛋白激酶信号途径的激活有密切关系 ,c fos调节MMP 1和TIMP 1和TIMP 2的含量 ,完成对愈合的调控。     

内皮抑素抑制膀胱癌皮下移植瘤生长及其机制的初步探讨

目的　探讨人内皮抑素对膀胱癌EJ细胞生长的作用及机制。　方法　 (1)体外实验观察内皮抑素对血管内皮细胞 (ECV30 4 )和膀胱癌EJ细胞生长的影响 ;(2 )EJ细胞移植入裸鼠皮下 ,观察内皮抑素对裸鼠皮下瘤生长情况的影响 ;(3)应用WesternBlot和RT PCR法观察皮下瘤组织中MMPs和TIMP 2的表达。　结果　 (1)内皮抑素具有抑制bFGF刺激后的内皮细胞ECV30 4及抑制EJ细胞增殖的作用 (P≤ 0 .0 5 ) ;(2 )治疗组皮下肿瘤质量 (0 .70± 0 .16 ) g ,对照组为 (1.14± 0 .2 1) g ,两组比较 ,差别有显著性意义 (P <0 .0 5 ) ;(3)人内皮抑素抑制裸鼠皮下膀胱癌瘤MMP 9mRNA表达 ,对MMP 2、TIMP 2、MT1 MMPmRNA表达无影响 ;(4)瘤组织中未检测到MMP 2、MMP 9和TIMP 2蛋白。　结论　人内皮抑素具有抑制膀胱肿瘤生长的作用 ,其作用机制与抑制MMP 9表达有关     

基质金属蛋白酶表达在膀胱癌侵袭转移中的作用

目的　探讨基质金属蛋白酶 1、2 (MMP 1,MMP 2 )在肿瘤侵袭转移过程中的作用。　方法　采用原位核酸分子杂交技术检测膀胱肿瘤手术标本中MMP 1、MMP 2mRNA的表达水平 ,结合临床资料进行分析。　结果　MMP 1和MMP 2mRNA在正常膀胱壁有少量表达。膀胱肿瘤中MMP 1和MMP 2mRNA的表达位于胞质内。MMP 1mRNA阳性率 4 3.5 % ,与细胞分级和临床分期无明显相关性。MMP 2mRNA阳性率 5 9.6 % ,其表达随细胞级别的升高而增强 (P <0 .0 5和P <0 .0 1) ,侵袭性癌与表浅性癌之间的表达差别有显著性意义 (P <0 .0 1)。　结论　MMP 2可作为判断肿瘤侵袭转移习性的指标 ,对临床制订治疗方案有一定意义。     

封闭负压引流技术对人慢性创面中胶原酶活性的影响

目的研究人慢性创面经封闭负压引流（vacuum—assisted closure，VAC）治疗前后，创面渗出液中胶原酶活性的变化，以部分阐明VAC促进慢性创面愈合的机理。方法取4例急性创面在术后1、2、3d的创面引流液（乳癌术后），同时收集6例慢性创面（4例静脉性溃疡，2例压力性溃疡）在VAC治疗前以及治疗后2、4、6d的创面渗出液，利用酶谱分析的方法，观察各时间点的渗出液对可溶性Ⅲ型胶原的降解情况，同时应用强力霉素抑制实验来分析渗出液中胶原酶的类型。结果急性创面引流液可以部分降解Ⅲ型胶原，随时间推移变化较小，慢性创面渗出液中的胶原酶活性较高，VAC治疗前基本将Ⅲ型胶原全部降解，随时间推移、降解减少，胶原酶活性下降，强力霉素抑制实验证明在100μmol／L浓度时无抑制，在600μmol／L浓度时出现部分抑制。结论在慢性创面渗出液中胶原酶活性增高，VAC的应用可以降低胶原酶的活性，阻止胶原蛋白大量降解，利于创面愈合，在慢性创面渗出液中胶原酶应主要是MMP-1型（成纤维细胞型）。     

Silver-impregnated vacuum-assisted closure in the treatment of recalcitrant venous stasis ulcers

Vacuum-assisted closure (VAC) has made a significant contribution to the treatment of acute and chronic wounds. Microdeformational forces from the VAC device accelerate granulation tissue formation when compared with moist saline dressing changes. We present 2 patients with multiple comorbid conditions and complex venous stasis ulcers that had persistent purulent drainage after conventional treatment modalities. Only after utilizing silver-impregnated VAC therapy (GranuFoam Silver), combining the antimicrobial benefits of silver with the advantages of VAC technology, were the wound beds adequately prepared for substantial split-thickness skin grafts. Based on these cases, the silver-impregnated VAC device may be a useful adjunct in wound bed preparation when standard therapies have failed to clear infected wounds. This may lead to improved healing rates and overall decreased wound burden in these complex patients.     

烧伤残余创面的序贯性治疗

目的寻找烧伤残余创面序贯性治疗的有效方法。方法选择烧伤残余创面患者25例，采取自身对照，分别以凡士林油纱（A组）、组织工程全层皮肤（HTAS，B组）覆盖创面，用含氧液冲洗、负压引流后覆盖HTAS（C组）。取创面标本检测细菌量、肿瘤坏死因子（TNF）α含量及基质金属蛋白酶（MMP）13mRNA表达水平，计算创面愈合率。结果治疗后第3、6、9、12天，患者C组创面细菌量分别为（5．30±1．60）、（1．30±0．80）、（1．70±0．60）和（0．60±0.10）集落形成单位（CFU）／ml，显著低于A、B组（P＜0．01）。C组创面经治疗后6d，TNF-α．含量和MMP-13mRNA表达水平分别为（0．650±0．040）ng／mg、0．210±0．010，明显低于A组[（1．550±0．370）ng／mg、1．040±0．050，P＜0．01]和B组[（0．810±0，080）ng／mg、0．640±0．030，P＜0．01]；B组肉芽组织TNF-α含量和MMP-13mRNA表达量显著低于A组（P＜0．01）。治疗后15、30d，C组创面愈合率明显高于A、B组（P＜0．01）。结论应用含氧液冲洗、负压引流后覆盖HTAS，可显著促进创面愈合，是一种有效的烧伤残余创面序贯性治疗方法。     

Microdeformational wound therapy: effects on angiogenesis and matrix metalloproteinases in chronic wounds of 3 debilitated patients

The vacuum-assisted closure (VAC) device causes microdeformations of the wound surface in contact with the foam. Because angiogenesis and matrix metalloproteinase (MMP) activity are altered in chronic wounds, we hypothesized that microdeformations stimulate capillary formation and affect MMP activity. A VAC device was used to deliver microdeformational wound therapy (MDWT) to the chronic wounds of 3 debilitated patients. Debrided tissue was obtained from wound areas with and without foam contact. Microvessel density and MMP activity were determined by immunohistochemistry and zymography, respectively. Microvessel density of MDWT-treated wounds was 4.5% (+/-0.8) compared with areas not covered by foam [1.6% (+/-0.1)] (P = 0.05) during the first week of treatment and 2.7% (+/-0.3) compared with untreated tissue [1.3% (+/-0.1)] (P = 0.03) during the second treatment week. Wounds subjected to MDWT had greater microvessel density compared with the same wound prior to treatment [1.5% (+/-0.3)] (P = 0.02). MMP-9/NGAL (neutrophil gelatinase-associated lipocalin), MMP-9, latent MMP-2, and active MMP-2 were reduced by 15%-76% in MDWT-treated wounds. MDWT provides a favorable wound-healing environment by increasing angiogenesis and decreasing MMP activity in chronic wounds.     

Vacuum assisted closure therapy for treatment of complex wounds in replanted extremities

The object of this study was to compare the outcomes of the vacuum assisted closure (VAC) therapy and conventional wound care with dressing change for treatment of complex wounds in patients with replantation of amputated upper and lower extremities. Data of 43 patients with replantation of amputated extremities from May 2004 to December 2011 were reviewed. There were 18 wounds of 18 patients with replantation, which were treated by dressing change and 26 wounds of 25 patients by VAC therapy. The outcomes were evaluated by the survival rate of replanted extremities, growth of granulation tissue, interval between wound treatment and secondary procedure and eventual secondary wound coverage methods. Vascular thromboses were found in 3 patients with wound treatment by dressing change and 5 by VAC. All replants of two groups of patients survived after salvage procedures. The wound score was 3.6 ± 0.7 in the conventional dressing change group and 5.8 ± 0.7 in the VAC group at the sixth day after treatment, respectively. The intervals between wound treatment and secondary wound coverage procedure were 12.0 ± 1.7 days in the dressing change group and 6.1 ± 0.7 days in the VAC group. Flaps were applied for wound coverage in 9 out of 18 (50.0%) wounds in the dressing change group and 5 out of 26 (19.2%) in the VAC group (P < 0.05), when the wounds of rest of patients were covered by the skin graft. The results showed that VAC could promote the growth of granulation tissue of wound, decrease the need of flap for wound coverage, and did not change the survival of replantation. © 2013 Wiley Periodicals, Inc. Microsurgery 33:620–624, 2013.     

封闭负压引流技术对兔耳急性创面愈合的影响

目的　研究封闭负压引流技术 (VAC)对急性创面愈合的影响 ,探讨其可能的治疗机制。方法　以免耳背急性全层皮肤缺损创面为模型 ,选取左侧耳背的创面为治疗组 ,给予封闭负压引流治疗 ;右侧创面为对照组 ,给予常规油纱敷料包扎治疗。分别在创面形成即刻和第 3、6、9天给创面摄影 ,应用图像分析软件计算创面愈合率。另外切取各时间点创面标本进行组织形态学观察。结果　在创面形成第 6天和第 9天 ,VAC组愈合率均高于对照组 ,两者差异有非常显著性 (P <0 .0 1)。组织形态学观察可见VAC组创面炎症反应较轻 ,肉芽组织生长和再上皮化速度均较对照组快。结论　封闭负压引流技术能减轻创面炎症反应 ,加快肉芽组织生长和再上皮化速度 ,显著促进兔耳背急性创面愈合。     

The use of vacuum-assisted closure therapy for the treatment of a large infected facial wound

Chronic wounds in difficult locations pose constant challenges to health care providers. Negative-pressure wound therapy is a relatively new treatment to promote wound healing. Laboratory and clinical studies have shown that the vacuum-assisted closure (VAC) therapy increases wound blood flow, granulation tissue formation, and decreases accumulation of fluid and bacteria. VAC therapy has been shown to hasten wound closure and formation of granulation tissue in a variety of settings. Accepted indications for VAC therapy include the infected sternum, open abdomen, chronic, nonhealing extremity wounds and decubitus ulcers. We report the first case of VAC therapy successfully used on a large infected wound to the face to promote healing.     

Effect of subatmospheric pressure on the acute healing wound

PURPOSE: Vacuum-assisted closure (VAC), originally developed as an adjunct to wound care, has gained popularity in managing complex, chronic wounds. This study was designed to compare VAC with traditional saline-wet-to-dry (WD) dressings on acute wound healing in a pig model. METHODS: Nine animals were divided into groups of 3. Three rows of 2, 4-cm diameter circular defects were excised on each animal. Vacuum-assisted closure therapy was applied to 2 adjacent wound beds, WD dressings were applied to 2 adjacent wound beds, and ventilated transparent dressing covered the 2 remaining wounds as controls. Random members from each group had their wounds harvested on postoperative days (POD) number 4, 7, and 9, respectively. The specimens were histopathologically evaluated and graded with regard to immature granulation tissue, mature granulation tissue, necropurulent surface crust, proliferating cell nuclear antigen (PCNA), and collagen deposition. RESULTS: The WD-treated wounds had less necropurulent material on the surface compared with the VAC and control groups (p < 0.05). Day 9 specimens demonstrated increased immature collagen in the VAC and WD groups compared with control. No other statistically significant variations existed between the treatment groups. CONCLUSIONS: Under the conditions of this study, the histopathologic observations do not support more rapid wound healing for the acutely injured VAC-treated wound compared with the WD-treated wound in young healthy pigs.