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。     

复发脑膜瘤血管内皮生长因子及增殖细胞核抗原的表达

目的探讨脑膜瘤细胞增殖能力、肿瘤复发与血管内皮生长因子（VEGF）蛋白表达之间的关系。方法应用免疫组织化学方法检测36例复发脑膜瘤及30例原发脑膜瘤标本的VEGF和增殖细胞核抗原（PCNA）表达。结果复发脑膜瘤VEGF蛋白阳性表达率89％（32／36）明显高于原发脑膜瘤43％（13／30）（X2=25．59，P〈0．01）。复发脑膜瘤PCNA指数（65．72±9．22）高于原发脑膜瘤（20．81±7．43，P〈0．05）。VEGF蛋白表达强阳性、弱阳性及阴性者的PCNA指数分别为78．64±10．02、49．45±8．31、6．23±1．45，差异有统计学意义（P〈0．01）。结论VEGF蛋白的表达水平与脑膜瘤的复发和增殖能力有关，VEGF蛋白表达水平可能是脑膜瘤复发的预测指标之一。     

特布他林对去甲肾上腺素和烧伤血清诱导大鼠神经胶质细胞血管内皮生长因子基因表达的影响

目的了解β2肾上腺素能受体激动剂特布他林对烧伤血清和去甲肾上腺素（NE）诱导神经胶质细胞血管内皮生长因子（VEGF）基因表达的影响。方法制备烧伤大鼠及正常大鼠血清。取出生1～3d乳鼠脑组织行原代神经胶质细胞培养，并在培养液中加入不同的刺激物，分为：（1）对照组：加入体积分数10％的正常大鼠血清；（2）NE1、NE2、NE3组：分别加入体积分数10％烧伤血清＋10、20、50μmol／LNE；（3）TBN1、TBN2、TBN3组：分别加入10、20、50μmol／L特布他林＋体积分数10％烧伤血清＋10、20、50μmol／LNE。采用蛋白质印迹法检测各组VEGF蛋白的表达；实时荧光定量PCR法检测VEGFmRNA的表达。结果对照组VEGF蛋白的表达处于较低水平，NE1、NE2、NE3组其表达随NE浓度的增大逐渐增加，而TBN1、TBN2、TBN3组的表达明显增加。对照组神经胶质细胞VEGF mRNA表达量[（5．72±0．12）拷贝数／g]较低，NEl、NE2、NE3组VEGF mRNA的表达[（13．26±0．03）、（10．37±0．04）、（14．87±0．55）拷贝数／g]均高于对照组（P〈0．01）。而TBN1、TBN2、TBN3组VEGF mRNA表达量[（13．39±0．19）、（15．77±0．11）、（16．00±0．07）拷贝数／g]也逐渐增加，除TBN1组外其余2组与各NE组比较，差异有统计学意义（P〈0．01）。结论特布他林可增强烧伤血清和NE诱导神经胶质细胞VEGF基因的表达。     

粘着斑激酶在PDGF-BB诱导MHCC-97H细胞黏附侵袭中的作用

目的观察不同浓度血小板衍生生长因子（PDGF-BB）诱导MHCC-97H细胞中粘着斑激酶（focal adhesion kinase，FAK）mRNA的动态变化及FAK在细胞黏附、侵袭的作用。方法不同浓度（1，2．5，5，10，25ng／m1）PDGF-BB诱导MHCC-97H人肝癌细胞，Real-time PCR方法检测FAK及MMP-2的mRNA动态变化，黏附实验、侵袭实验分别检测诱导后MHCC-97H细胞的黏附、侵袭能力的变化。结果诱导后MHCC-97H细胞FAKmRNA水平上调，在10ng／ml浓度时达到最高，为对照组的112．6倍；MMP-2mRNA水平上调并与FAKmRNA水平上调呈一致性，在10ng／ml浓度时达到最高，为对照组的56．9倍。诱导后各组黏附率分别为（66±1．84）%、（69±1．41）％、（69±2．42）％、（71±1．37）%和（66±3．28）%，与对照组的（54±2．08）％比较均有统计学意义（P〈0．001）；诱导后5ng／ml和10ng／ml组侵袭细胞数分别为（26．63±4．5）、（28．75±4．2）个，与对照组（21．5±4．0）个比较有显著性差异（P〈0．05，P〈0．001）。诱导后黏附率与侵袭细胞数变化在10ng／ml浓度时都达到高峰。结论PDGF-BB上调MHCC-97H细胞中FAK表达，上调FAK促进MHCC-97H细胞的黏附和侵袭能力。     

冠状动脉旁路移植手术方式对甲状腺激素代谢的影响

目的观察体外循环（on-pump）心脏停跳下和非体外循环（off-pump）心脏不停跳冠状动脉旁路移植手术围术期甲状腺激素的变化规律。方法冠脉旁路手术病人40例，分为Off-pump组和Onpump组，每组20例，分别测定围术期甲状腺功能指标。结果术前两组病例甲状腺功能指标均在正常值范围内。Off-pump组T3和FT3术后显著下降，术后第1d降至最低[（T3：（39．7±6．2）ng，dl对（78．6±12．1）mg/dl，P〈0．01；FT3（1．6±0．5）pg/ml对（2．4±0．6）pg/ml，P〈0．05]；T4在正常值范围内变化，术后第1d降至最低[（5．3±0．8）μg/dl对（7．2±0．7）μg/dl，P〈0．05]；FT4在正常值范围内波动，各阶段检测值与术前差异无统计学意义（P〉0．05）；rT3术后逐渐升高，术后第3d达高峰[（82．1±26．8）ng/dl对（42．3±15．6）ng/dl，P〈0．01]；TSH术中逐渐下降，术后第1d降至最低[（0．4±0．2）tdU／ml对（1．3±0．6）μIU／ml，P〈0．01]，术后第7d恢复术前水平。围术期两组病例甲状腺功能指标变化趋势相似，差异无统计学意义（P〉0．05）。结论On-pump与Off-pump下冠状动脉旁路移植手术均影响甲状腺激素代谢，两组病人术后均存在低B综合征。     

胃减容术3种机械通气对患者氧合情况的比较

目的：探讨病态肥胖患者行腹腔镜胃减容术时，3种机械通气参数对患者氧合情况的影响。方法：选择行腹腔镜胃减容术的病态肥胖24例，按手术时间顺序分为3组，每组8例。A组：潮气量12ml／kg，呼吸频率10次／min；B组：潮气量20ml／kg，呼吸频率10次／min；C组：潮气量12ml／kg，呼吸频率20次／min。分别于气腹前（T1）、气腹后（T2）抽取动脉血测量pH、PO2、PCO2、Plat（平台压）、Peak（峰压）、AaDO2（肺泡-动脉氧分压差）。结果：气腹后B组Plat（33．2±1．8）cmH2O、Peak（36．3±1．6）cmH2O显著高于A组Plat（29．5±3．9）cmH2O（q=3．053，P〈0．05）、Peak（33．0±2．9）cmH2O（q：2．823，P〈0．05）和C组Plat（28．0±4，1）cmH2O（q：4．290，P〈0．05）、Peak（31．8±2．8）cmH2O（q：4．704，P〈0．05）。其他观察指标无统计学差异。结论：病态肥胖患者在腹腔镜下施行胃减容术时，加大潮气量20ml／kg或加快呼吸频率20次／min不能改善患者的氧合情况。     

自制多器官保存液对肾皮质能量代谢的影响

目的寻找理想的器官保存液，以提高器官移植的长期疗效。方法应用高效液相系统（HPLC），检测自制多器官保存液（SMO液）和对照组（HC-A液和UW液）对犬肾低温保存中皮质腺苷酸含量改变的影响。取皮质标本匀浆、沉淀蛋白、离心，HPLC检测上清中腺苷酸含量，根据标准曲线方程和加样回收率计算ATP、ADP、AMP含量、比值和能荷（EC）。结果随保存时间延长，各组皮质腺苷酸含量下降；保存12、36、72h后SMO组皮质ATP含量（3．7±0．5、3．1±0．3、2．3±0．5nmol／mg蛋白）高于HC—A组（2．8±0．2、2．0±0．2、1．5±0．2nmol／mg蛋白），P〈0．05，SMO组EC（0．38±0．04、0．37±0．05、0．35±0．04）高于HCfA组（0．32±0．03、0．30±0．04、0．28±0．03），P〈0．05，在多数保存时点SMO组ADP、ATP／AMP高于HC—A组（P〈0．05），但2组间AMP、ATP／ADP比值差异无统计学意义（P〉0．05）；犬肾保存期间SMO组腺苷酸含量及EC与UW组比较差异无统计学意义（P〉0．05）。结论低温保存期间SMO液能够维持皮质内较高的ATP和EC水平，减轻肾脏低温缺血损伤。     

自体骨髓细胞经门静脉移植治疗肝硬化与肝功能不全的临床研究

目的观察自体骨髓细胞经门静脉移植对肝硬化和肝功能不全的治疗效果。方法2005年2月至2006年6月在我科接受手术治疗的40例肝硬化门静脉高压症患者（脾切除、断流术或内镜食道曲张静脉套扎术）,被随机分为治疗组和对照组,每组20例。两组患者于术中埋置“门静脉导管-皮下药盒”,术后3-4周,治疗组经移植通道输注自体骨髓细胞,而对照组只输注生理盐水。在第1次输注后每隔1个月再重复进行输注,共输注3次。第3次输注后1个月进行疗效评价。结果（1）两组恢复均顺利,未发现与移植操作有关的不良反应或并发症。（2）丙氨酸转氨酶、总胆红素、白蛋白和凝血酶原时间：治疗组分别由（60±52）μmol／L、（26±15）μmol／L、（33±5）μmol／L和（18±2）s变为（26±15）μmol／L、（14±8）μmol／L、（41±3）μmol／L和（12±2）s（P〈0．01）;对照组分别由（47±37）μmol／L、（22±23）μmol／L、（35±4）μmol／L和（18±4）s变为（65±51）μmol／L、（19±42）μmol／L、（35±4）μmol／L和（18±4）s（P〉0．05）;治疗组优于对照组（P〈0．01）。（3）血清透明质酸和前胶原Ⅲ肽：治疗组分别由（188±160）ng／ml和（13±18）ng／ml变为（104±80）ng／ml和（8±9） ng／ml（P〈0．05）;对照组分别由（79±193）ng／ml和（10±16）ng／ml变为（136±187）ng／ml和（9±17）ng／ml（P〉0．05）;治疗组亦优于对照组（P〈0．01）。结论自体骨髓细胞经门静脉移植可改善肝功能和肝纤维化血清学指标。     

转染人Endostatin基因对胰腺癌SW1990细胞增殖、凋亡及血管内皮生长因子表达的影响

目的观察转染人Endostatin（hEndostatin）基因对胰腺癌细胞株SW1990增殖、凋亡及血管内皮生长因子（VEGF）表达的影响。方法携带hEndostatin基因的复制缺陷型重组腺病毒载体体外转染SW1990细胞，MTT法检测细胞增殖，流式细胞仪检测细胞凋亡，实时荧光定量PCR和ELISA分析检测hEndostatin和VEGF的表达。建立裸鼠胰腺癌模型，随机分为3组（n=8），瘤体内分别注射磷酸盐缓冲液（PBS组）、报告基因LacZ重组腺病毒（Ad—LacZ组）、hEndostatin重组腺病毒（Ad—bEnd组）100山，隔日1次，共4次。4周后免疫组织化学染色检测肿瘤组织中hEn．dostatin、VEGF、增殖性细胞核抗原（PCNA）的表达，TUNEL法检测肿瘤细胞凋亡。结果体外转染hEndostatin基因不影响SW1990细胞的增殖和凋亡（P〉0．05），但下调VEGF的表达（P〈0．01），下调作用第5天最大，在mRNA和蛋白水平的抑制率分别为84．67％和81．41％。体内转染4周后，Ad．bend组VEGF、PCNA阳性表达率分别为（36．3±7．1）％、（38．2±3．9）％，显著低于Ad．LacZ组的（81．2±6．6）％、（93．2±4．9）％和PBS组的（78．4±6．2）％、（90．1±5．7）％（P〈0．01）；肿瘤细胞凋亡率为（31．2±5．4）％，显著高于Ad—LacZ组的（9．4±4．9）％和PBS组的（8．5±3．7）％（P〈0．01）。结论hEndostatin基因转染抑制SW1990细胞的体内增殖并促进凋亡；下调SW1990细胞内源性VEGF的表达可能是抗肿瘤的作用机制之一。     

合成短肽S247对呼吸机所致肺损伤中肺上皮细胞凋亡的影响

目的 研究合成短肽S247对呼吸机相关性肺损伤凋亡的影响，探索其在肺损伤中的防护作用。方法 30只清洁级雄性SD大鼠随机分为3组：对照组（C组）、肺损伤组（Ⅰ组）和S247治疗组（S组），每组10只，大潮气量损伤性通气4h建立呼吸机相关性肺损伤模型（呼吸参数：Vt=40ml／kg体重，呼吸频率=80次／min，FiO2=21％），光镜下观察肺病理改变，测定肺湿／干比；收集肺泡灌洗液（BALF），考马思亮兰法测BALF中总蛋白含量，光境下行WBC计数，免疫组织化学检查肺组织Fas、Caspase-3蛋白的表达水平，TUNEL法检测肺组织细胞凋亡情况。结果 Ⅰ组肺病理改变明显，各组肺湿／干比、BALF中总蛋白含量、WBC计数差异都有统计学意义（P〈0．05或P〈0．01），Ⅰ组Fas、Caspase-3蛋白的表达水平（0．359±0．055；0．152±0．007）显著增高（P〈0．01），凋亡指数（60．000±7．461）明显增加（P〈0．01）；在用S247后，Fas、Caspase-3蛋白的表达水平（0．240±0．062；0．146±0．004）显著下降（P〈0．05），凋亡指数（47．857±3．716）有所减少（P〈0．05）。结论 S247能通过抑制细胞凋亡，有效降低呼吸机相关性肺损伤的程度。     

Comparison of the postburn hyperdynamic state and changes in lung function (effect of wound bacterial content)

The pulmonary and systemic response to a full-thickness burn (15% of total body surface area) was determined in 15 adult sheep. Also compared was the effect of wound bacterial content and prostanoid release on this response. Burn wound thromboxane A2, measured as TxB2, and prostacyclin, measured as 6-keto-PGF1 alpha, were measured in burn wound lymph. Animals were monitored for 7 days. On the final day, a full-thickness biopsy specimen of burn tissue was obtained for quantitative bacteriology. Wounds with 10(4) or less organisms per gram of burn tissue were considered colonized, whereas those with 10(5) or more organisms per gram of burn tissue indicated wound infection. Seven sheep had 10(4) or less bacteria and the remaining eight sheep had 10(6) or greater bacteria. We noted a significant mean increase in cardiac index from a baseline of 5 to 6.2 L/min/m2, a decrease in systemic vascular resistance from 16 to 12 mm Hg/L/min, and a mean increase in oxygen consumption from a baseline of 135 to 165 ml/min/m2 during the 7-day study period. There were no differences in these responses between the colonized and the infected wounds. Pulmonary artery pressure increased from a mean baseline of 19 to 24 mm Hg and arterial oxygen tension (PaO2) decreased from a baseline of 90 to 80 mm Hg in the infected wound group, with values remaining at baseline in the colonized wound group. These changes corresponded with an increase in lymph and plasma TxB2 from a baseline of 200 to 210 pg/ml to 1000 +/- 250 and 600 +/- 190 pg/ml, respectively. Values in the animals with colonized wounds were not significantly increased.(ABSTRACT TRUNCATED AT 250 WORDS)     

糖尿病足溃疡患者创面局部注射胰岛素对全身血糖及创面的影响

目的 观察糖尿病足溃疡患者创面局部注射胰岛素对全身血糖和创面肉芽组织形成的影响. 方法 选择2009年6月-2010年6月在笔者单位住院治疗的糖尿病足患者32例,按随机数字表法分为胰岛素组和对照组,每组16例.所有患者预先清创.(1)胰岛素组:将胰岛素计算剂量的1/2用生理盐水稀释成1 mL后,浸润注射于溃疡创面基底部,另1/2计算量按常规行腹部皮下注射,均为2次/d.(2)对照组:将胰岛素计算剂量的全量常规注射于腹部皮下,溃疡创面基底部浸润注射1 mL生理盐水,均为2次/d.2组患者均连续注射7d.于患者每次注射前及注射后0.5、1.0、2.0、4.0 h,分别检测其空腹血糖值.于首次注射前及注射3、5、7 d,评估创面肉芽组织生长程度;取创面组织标本观察CD34表达情况,据此计算创面微血管密度(MVD).对实验数据行t检验.结果 2组患者观察期内空腹血糖值维持在6.6～12.8(10.0±2.2)mmol/L,各时相点组间比较,差异均无统计学意义(t值为0.000～2.209,P值均大于0.05).注射第5天起,胰岛素组患者局部创面肉芽组织明显增多;第7天达生长高峰[(59.06±1.58％],与对照组[(23.61±1.57)％]比较,差异有统计学意义(t=17.420,P=0.0.000).CD34表达情况显示,胰岛素组注射3 d起有新生血管形成,但此时MVD与对照组接近(t=0.0247,P＞0.05);注射5、7d,胰岛素组创面组织MVD每200倍视野中分别为(8.34±0.48)、(11.22±0.97)个,明显多于对照组的(4.42±0.14)、(5.44±1.13)个,t值分别为16.568、27.664,P值均小于0.01. 结论 糖尿病足溃疡患者创面局部注射胰岛素对全身血糖有较明显影响,能加速肉芽组织生长,促进创面愈合.     

Recombinant human erythropoietin stimulates angiogenesis and wound healing in the genetically diabetic mouse

The effects of recombinant human erythropoietin (rHuEPO) in diabetes-related healing defects were investigated by using an incisional skin-wound model produced on the back of female diabetic C57BL/KsJ-m(+/+)Lept(db) mice (db(+)/db(+)) and their normoglycemic littermates (db(+/+)m). Animals were treated with rHuEPO (400 units/kg in 100 microl s.c.) or its vehicle alone (100 microl). Mice were killed on different days (3, 6, and 12 days after skin injury) for measurement of vascular endothelial growth factor (VEGF) mRNA expression and protein synthesis, for monitoring angiogenesis by CD31 expression, and for evaluating histological changes. Furthermore, we evaluated wound-breaking strength at day 12. At day 6, rHuEPO injection in diabetic mice resulted in an increase in VEGF mRNA expression (vehicle = 0.33 +/- 0.1 relative amount of mRNA; rHuEPO = 0.9 +/- 0.09 relative amount of mRNA; P < 0.05) and protein wound content (vehicle = 23 +/- 5 pg/wound; rHuEPO = 92 +/- 12 pg/wound; P < 0.05) and caused a marked increase in CD31 gene expression (vehicle = 0.18 +/- 0.05 relative amount of mRNA; rHuEPO = 0.98 +/- 0.21 relative amount of mRNA; P < 0.05) and protein synthesis. Furthermore, rHuEPO injection improved the impaired wound healing and, at day 12, increased the wound-breaking strength in diabetic mice (vehicle = 12 +/- 2 g/mm; rHuEPO 21 +/- 5 g/mm; P < 0.05). Erythropoietin may have a potential application in diabetes-related wound disorders.     

Aminated β-1,3-d-glucan improves wound healing in diabetic db/db mice

Delayed wound healing in diabetes is caused by neuropathy, vascular changes, and impaired cellular response to the injury. Macrophages are crucial in normal wound healing, and impaired functions of these cells have been shown in diabetes. beta-1,3-D-glucans stimulate macrophage function. This open-label study was performed to see if aminated beta-1,3-D-glucan (AG) stimulates wound healing in diabetes. Four groups (1-4) of diabetic db/db mice and one nondiabetic control group, db/+(5) were studied: group 1 (n=11): topical AG; group 2 (n=10): topical AG and subcutaneous insulin; group 3 (n=14): topical placebo and subcutaneous insulin; group 4 (n=10): diabetic control (placebo); group 5 (n=12): normal control (placebo). At the end of the experiments fasting blood glucose and A1C were (mean +/- SE) as follows: Group 1: 30.5 +/- 1.9 mmol/L and 11.3 +/- 0.6%; group 2: 12.0 +/- 1.7 mmol/L and 8.0 +/- 0.6%; group 3: 15.4 +/- 2.4 mmol/L and 7.4 +/- 0.3%; group 4: 32.6 +/- 2.6 mmol/L and 12.3 +/- 0.6%; group 5: 7.2 +/- 0.4 mmol/L and 3.9 +/- 0.04%, respectively. The closed wound area was the same in group 1 (AG alone) and group 2 (AG plus insulin) after 17 days, 57.3 +/- 4.7 vs. 50.1 +/- 4.9% (p=0.7).The results of these two groups were superior to group 3 (insulin treatment alone, 32.0 +/- 4.3%, p<0.001) and diabetic controls (38.2 +/- 5.1%, p=0.001). The macrophage-stimulant AG improves wound healing in db/db mice.     

Topical ibuprofen decreases thromboxane release from the endotoxin-stimulated burn wound

A full-thickness burn wound in adult sheep releases prostanoids when they are injected locally with E. coli endotoxin, 2 micrograms/kg, resulting in an increase in pulmonary artery pressure (Ppa) from 20 +/- 3 to 34 +/- 5 mm Hg, and a decrease in mean arterial oxygen tension (PaO2) from 88 +/- 6 to 70 +/- 5 torr; this corresponds to an increase in venous plasma TxB2 content from a baseline of 220 +/- 79 pg/ml to 440 +/- 90 pg/ml. Burn prostanoid production, measured in lymph, increased ten- to fifteen-fold for both thromboxane A2, measured as TxB2, and prostacyclin, 6-keto-PGF1 alpha. The intravenous administration of ibuprofen, 12.5 mg/kg, eliminated both the increase in Ppa and decrease in PaO2 as well as the increase in burn lymph prostanoids. However, plasma prostanoids were also decreased below baseline, a potentially deleterious effect. A topical ibuprofen cream, 5% ibuprofen in a water-soluble ester, was applied to the burn hide every 6 hrs x 4 after which endotoxin was again injected below the hide. The pulmonary dysfunction was prevented as was the increase in plasma TxB2 with the value remaining at baseline. Burn lymph levels were only increased three- to five fold. Ibuprofen levels in burn lymph were maintained at 1-2 mcg/ml. The addition of the cream to the burn, however, did increase the wound bacterial content to 10(5)-10(7) bacteria/gram of tissue compared to 10(2)-10(3) for the dry, untreated burn, probably due to softening of the burn. Topically applied ibuprofen, therefore, can decrease burn wound prostanoid production from local endotoxin, preventing lung dysfunction.(ABSTRACT TRUNCATED AT 250 WORDS)     

部分创面外用抗菌药物与成纤维细胞生长因子2、表皮生长因子、重组人生长激素对成纤维细胞生物学特性影响的实验研究

目的观察部分创面外用抗菌药物与成纤维细胞生长因子(FGF)2、表皮生长因子 (EGF)、重组人生长激素(rhGH)对成纤维细胞生物学特性的影响。方法体外培养成纤维细胞, 按所加药物不同分为对照组(常规培养)、丁胺卡那霉素(0．021、0．210、2．100 mg/L)组、庆大霉素(5、 50、500 mg/L)组、氯霉素(0．01、0．10、1．00 mg/L)组、磺胺米隆(5、10 g/L)组、FGF2(2 400 U/ml)组、 EGF(2 000 U/ml)组及rhGH(0．016、0．160、1．600 g/L)组。用噻唑蓝(MTT)法测定各组成纤维细胞增殖活性[吸光度(A)值],用流式细胞仪检测细胞周期,并于显微镜下观察细胞形态的变化。结果 (1)MTT法检测:与对照组A值0．455 3±0．021 7比较,各种剂量丁胺卡那霉素组、庆大霉素组、氯霉素组、磺胺米隆组成纤维细胞A值均明显降低(P<0．05或0．01),其中磺胺米隆(5、10 g/L)组降低最明显,分别为0．101 3±0．001 1、0．095 0±0．004 1(P<0．01)。FGF2组及0．016 g/L rhGH 组细胞A值明显高于对照组(P<0．05),而EGF组及0.160、1．600 g/L rhGH组A值与对照组接近 (P>0．05)。(2)细胞周期检测:对照组细胞增殖指数(PI)为(9．63±0．45)％,与之比较,0．210 mg/L丁胺卡那霉素组细胞PI值无明显变化(P>0．05),FGF2组、EGF组及0．016 g/L rhGH组PI值均明显升高,分别为(46．76±2．33)％、(42．30±1．41)％、(13．29±0．47)％(P<0．05或0．01)。 (3)形态学观察:对照组、EGF组及0．160、1．600 g/L rhGH组成纤维细胞数目较多,呈长条形或梭形, 轮廓不清,透明度高;丁胺卡那霉素组、庆大霉素组、氯霉素组、磺胺米隆组细胞数目较少,形态不规则,轮廓清晰,透明度低,细胞内多有颗粒样物质及空泡;FGF2组、0．016 g/L rhGH组细胞分布均匀、密集,呈长条形或梭形,核分裂相多见,轮廓不清,透明度高。结论不同创面外用药物对成纤维细胞生物学特性的影响各异,在创面修复过程中应选择合适的创面外用药物,以促进愈合并抑制瘢痕增生。     

LPS-pretreatment adipose-derived mesenchymal stromal cells promote wound healing in diabetic rats by improving angiogenesis

Mesenchymal stem cells (MSCs) play a key role in wound healing, and the advantages of pretreated MSCs in wound healing have previously been reported. In the present study, we investigated the impact of LPS pretreated human adipose-derived MSCs on skin wound healing in diabetic rats. We found that some improvements occurred through improving angiogenesis. Then, we scrutinized the impact of lipopolysaccharide (LPS) treatment on human adipose-derived MSCs in a high-glucose (HG) medium, as an in vitro diabetic model. In vivo findings revealed significant improvements in epithelialization and angiogenesis of diabetic wounds which received LPS pre-MSCs. Particularly, LPS pre-MSCs-treated diabetic wounds reached considerably higher percentages of wound closure. Also, the granulation tissue of these wounds had higher pronounced epithelialization and more vascularization compared with PBS-treated and MSCs-treated diabetic ones by CD31, VEGF, CD90, collagen 1, and collagen 3 immunostaining. Western-blots analyses indicated that LPS pre-MSCs led to the upregulation of vascular endothelial growth factor (VEGF) and DNMT1. In addition, significantly higher cell viability (proliferation/colonie), and elevated VEGF and DNMT1 protein expression were observed when MSCs were treated with LPS (10 ng/ml, 6 h) in HG culture media. Based on these findings, it is suggested that LPS pre-MSCs could promote wound repair and skin regeneration, in some major processes, via the improvement of cellular behaviors of MSCs in the diabetic microenvironment. The beneficial advantages of LPS treated with mesenchymal stem cells on wound healing may lead to establishing a novel approach as an alternative therapeutic procedure to cure chronic wounds in diabetic conditions.     

明胶海绵微粒化疗栓塞肝细胞癌后外周血中血管内皮生长因子水平变化

目的通过观察肝细胞癌（HCC）患者外周血中血管内皮生长因子（VEGF）水平在GSPs化疗栓塞治疗前后的变化,探讨该方法对肿瘤血管生成的影响。方法应用ELISA双抗体夹心法检测30例原发性HCC患者（HCC组）GSPs化疗栓塞术前1天、术后4、7及28天和30名健康志愿者（对照组）外周血中VEGF水平。结果 HCC患者术前外周血中VEGF含量明显高于对照组（P〈0.05）。HCC患者术后4天[（232.56±119.40）pg/ml]、7天[（290.33±97.17）pg/ml]外周血中VEGF水平较术前1天[（106.79±55.85）pg/ml]明显增高（P均〈0.05）;术后28天[（233.54±106.60）pg/ml]低于术后7天[（290.33±97.17）pg/ml],但差异无统计学意义。结论 GSPs化疗栓塞治疗HCC术后VEGF规律性的变化可反映肿瘤血管生成状态,为术后抗血管内皮生长治疗提供理论依据。     

成纤维细胞生长因子在烧伤创面内变化的临床观察

目的 探讨临床烧伤创面愈合过程中成纤维细胞生长因子（ＦＧＦ）的变化及其作用。方法采用免疫组织化学方法,对烧伤创面肉芽中心部、肉芽与愈合皮肤的交界部、愈合皮肤和自身正常皮肤,以及Ⅲ度烧伤创面不同部位和Ⅱ度烧伤创面愈合过程中ＦＧＦ表达变化进行了观察。结果 Ⅲ度烧伤创面不同部位ＦＧＦ表达强度不同,以创面的肉芽中心部ＦＧＦ含量最多,肉芽与愈合皮肤的交界部ＦＧＦ含量次之,愈合皮肤ＦＧＦ含量较少;每个创面ＦＧ