荷负电气溶胶治疗对大鼠烫伤创面白细胞介素-8、10表达的影响及意义

Objective To discuss the influence of aerosol bioelectricity on the expression of interleukin (IL) -8 and IL-10 in wound healing of burned rats. Methods The deep Ⅱ degree scalding models were established in Sprague Dawley (SD) rats. Rats were randomly divided into experimental group (n1 =20) and control group (n2 =20). The rats in experimental group were treated with aerosol bioelectricity.Samples were collected at the first to eleventh day post-scalding. Immunohistochemistry and image analysis methods were conducted to examine the expression of IL-8 and IL-10 in both experimental and control groups. Results The average wound healing time in experimental group was 7. 00 ± 1. 15 days, and that in control group was 9. 00 ± 1. 34 days. IL-8 and IL-10 were observed mainly in polylmorphonuclear and mononuclear cells in both experimental and control groups on the 1 st day. On the third day, fibroblasts abounded, IL-8 expression was increased evidently and reached a peak. The peak value (6. 73 ± 1. 36) in experimental group was lower significantly than that in control group ( 2. 85 ± 0. 72, P ＜ 0. 01). From the 5th to 11th day, IL-8 expression was declined rapidly. IL-10 was expressed in keratode cells and had the peak value in experimental group (1. 24 ±0. 15) and control group (5. 69 ± 1. 32) on the 3rd day. IL-10 expression was declined gradually from the 5th to 11th days. The expression level of IL-10 in experimental group was significantly higher than in control group from the 3rd day to 11th days post-scalding (P＜0. 01). On the 3rd day, both IL-8 and IL-10 in experimental and control groups were expressed abundantly , and there was negative relationship between them (r = - 0. 862, P ＜ 0. 01). Conclusion Aerosol bioelectricity can indicate active cells proliferation through down-regulating the expression of IL-8 and up-regulating the expression of IL-10, accelerating burned wound healing.     

荷负电气溶胶治疗对大鼠烫伤创面白细胞介素-8、10表达的影响及意义

Objective To discuss the influence of aerosol bioelectricity on the expression of interleukin (IL) -8 and IL-10 in wound healing of burned rats. Methods The deep Ⅱ degree scalding models were established in Sprague Dawley (SD) rats. Rats were randomly divided into experimental group (n1 =20) and control group (n2 =20). The rats in experimental group were treated with aerosol bioelectricity.Samples were collected at the first to eleventh day post-scalding. Immunohistochemistry and image analysis methods were conducted to examine the expression of IL-8 and IL-10 in both experimental and control groups. Results The average wound healing time in experimental group was 7. 00 ± 1. 15 days, and that in control group was 9. 00 ± 1. 34 days. IL-8 and IL-10 were observed mainly in polylmorphonuclear and mononuclear cells in both experimental and control groups on the 1 st day. On the third day, fibroblasts abounded, IL-8 expression was increased evidently and reached a peak. The peak value (6. 73 ± 1. 36) in experimental group was lower significantly than that in control group ( 2. 85 ± 0. 72, P ＜ 0. 01). From the 5th to 11th day, IL-8 expression was declined rapidly. IL-10 was expressed in keratode cells and had the peak value in experimental group (1. 24 ±0. 15) and control group (5. 69 ± 1. 32) on the 3rd day. IL-10 expression was declined gradually from the 5th to 11th days. The expression level of IL-10 in experimental group was significantly higher than in control group from the 3rd day to 11th days post-scalding (P＜0. 01). On the 3rd day, both IL-8 and IL-10 in experimental and control groups were expressed abundantly , and there was negative relationship between them (r = - 0. 862, P ＜ 0. 01). Conclusion Aerosol bioelectricity can indicate active cells proliferation through down-regulating the expression of IL-8 and up-regulating the expression of IL-10, accelerating burned wound healing.     

铜绿假单胞菌感染大鼠皮肤创面愈合过程中TGF-β1及胶原蛋白含量的变化

目的 探讨大鼠慢性皮肤溃疡创面感染铜绿假单胞菌后,TGF-β1和胶原Ⅰ、Ⅲ蛋白表达的变化.方法 24只8周龄雌性Wister大鼠随机分为单纯创面组(A组)和创面+铜绿假单胞菌接种组(B组).分别在术后第1、3、7、10天观察创面上皮化率、收缩率及中性粒细胞情况;并采用ELISA方法测定创面第1、3、7、10天TGF-β1和胶原Ⅰ、Ⅲ蛋白的表达情况.结果 A组上皮化率在第7天高于B组,收缩率低于B组.随着时间的延长,A组中性粒细胞在第3天增加到最多,随后逐渐减少,而B组中性粒细胞第1天达到最多.2组TGF-β1表达在术后呈上升趋势,B组TGF-β1在第3天降低,随着时间延长回升,第7天2组比较差异有统计学意义(P＜0.05).胶原Ⅰ、Ⅲ蛋白表达随着时间的延长呈下降趋势,B组胶原Ⅲ蛋白表达在第7、10天差异有统计学意义(P＜0.05).A组第1、3天胶原Ⅰ、Ⅲ蛋白表达高于B组,在第7、10天则低于B组,且胶原Ⅲ蛋白在第3天显示A组高于B组,差异有统计学意义(P＜0.05).结论 皮肤溃疡创面感染铜绿假单胞菌后延迟了TGF-β1和胶原Ⅰ、Ⅲ蛋白的表达,可能影响创面的正常愈合.

Abstract：

Objective To explore the different expression of TGF-β1 and collagen during the healing process of wound infected by pseudomonas aeruginosa(PAO1).Methods 24 female Wister rats were randomly divided into pure wound group(group A)and wound+PAO1 group(group B).The reepithelial rate,shrinkage rate and neutrophils number on the wounds were observed on the 1st,3rd,7th and 10th day after operation.The expression of TGF-β1 and collage Ⅰ,Ⅲ was also detected.Results On the 7th day,the re-epithelial rate in group A was higher than that in group B,while the shrinkage rate in group A was lower than that in group B.The neutrophils number increased to peak on the 1 st day in group B,but on the 3rd day in group A.The TGF-β1 expression increased after operation in both groups,but it decreased in group B on the 3rd day and re-increased after that.The TGF-β1 expression was significantly different between the two groups on the 7th day(P＜0.05).The expression of collagen Ⅰ and Ⅲ decreased during healing.The expression of collagen Ⅲ in group A was higher on the 3rd day and was lower on the 7th and 10th day than that in group B,showing a significant difference(P＜0.05).Conclusions PAO1 infection could delay the expression of TGF-β1 and collagen Ⅰ,Ⅲon wound,which may interfere the healing process of wound.     

完全去交感肾上腺素能神经支配对小鼠前列腺癌生物学行为的影响

Objective To investigate the effects of complete denervation of sympathetic adrenergic nerves on the biological behaviors of prostate cancer in C57BL/6 mice. Methods Fifty male C57BL/6 mice, 10 weeks old, were randomly divided into two groups. By using a micro-syringe, RM-1 cells (0. 5 ×106) were injected into bilateral dorsal prostate capsules of mice, at the same time, bilateral hypogastric sympathetic nerves of the mice were cut (experimental group) or not (control group). At the sixth day postoperation, 5 mice were sacrificed in each group every three days to observe the local growth, invasion and metastasis of prostate cancer to pelvic nodes and other organs. An immunohistochemical determination of the hypogastric nerves was made by using an antibody against tyrosine hydroxylase ( TH) , a marker for sympathetic nerves. At the 15th day, 10 mice left were fed to death to calculate the life span of tumor-bearing mice. Results The volume of the prostate was gradually increased and confirmed as prostate cancer histologically. The prostate volume in experimental group was (0. 034 ± 0. 008) , (0. 339 ±0. 040) , (0. 829 ±0. 090) , ( 1. 169 ±0. 093) cm3 on the day 6, 9, 12, 15 after operation, respectively,while that in control group was (0.034 ± 0. 008), (0. 316 ± 0. 050), (0. 824 ± 0.071), (1. 236 ±0. 103) cm3, respectively (all P ＞ 0. 05). At the 12th day, muscle tissues around the prostate invasion by prostate cancer were observed in 3 cases of experimental group and 4 cases of control group; at the 15th day, invasion of seminal vesicles and bladder was found in 4 cases of experimental group and 4 cases of control group. Three and 4 mice had metastasis of pelvic nodes at 12th day and 15th day respectively in the experimental group, and 4 and 4 respectively in control group. At the 15th day post operation, in control group, liver parenchyma metastasis and renal pelvis metastasis were observed in 1 and 1 case, respectively, while in experimental group, no distant metastasis was observed. The life span of tumor-bearing mice in the experimental and control groups was (15.80±0.84) and (16.00±0.71) days, respectively (P＞0. 05 ) . Conclusion Complete denervation of sympathetic adrenergic nerves seems to have no significant effects on the local growth, invasion and pelvic nodes metastasis of prostate cancer in C57BL/6 mice, but may have some inhibition effects on its distant metastasis.     

完全去交感肾上腺素能神经支配对小鼠前列腺癌生物学行为的影响

Objective To investigate the effects of complete denervation of sympathetic adrenergic nerves on the biological behaviors of prostate cancer in C57BL/6 mice. Methods Fifty male C57BL/6 mice, 10 weeks old, were randomly divided into two groups. By using a micro-syringe, RM-1 cells (0. 5 ×106) were injected into bilateral dorsal prostate capsules of mice, at the same time, bilateral hypogastric sympathetic nerves of the mice were cut (experimental group) or not (control group). At the sixth day postoperation, 5 mice were sacrificed in each group every three days to observe the local growth, invasion and metastasis of prostate cancer to pelvic nodes and other organs. An immunohistochemical determination of the hypogastric nerves was made by using an antibody against tyrosine hydroxylase ( TH) , a marker for sympathetic nerves. At the 15th day, 10 mice left were fed to death to calculate the life span of tumor-bearing mice. Results The volume of the prostate was gradually increased and confirmed as prostate cancer histologically. The prostate volume in experimental group was (0. 034 ± 0. 008) , (0. 339 ±0. 040) , (0. 829 ±0. 090) , ( 1. 169 ±0. 093) cm3 on the day 6, 9, 12, 15 after operation, respectively,while that in control group was (0.034 ± 0. 008), (0. 316 ± 0. 050), (0. 824 ± 0.071), (1. 236 ±0. 103) cm3, respectively (all P ＞ 0. 05). At the 12th day, muscle tissues around the prostate invasion by prostate cancer were observed in 3 cases of experimental group and 4 cases of control group; at the 15th day, invasion of seminal vesicles and bladder was found in 4 cases of experimental group and 4 cases of control group. Three and 4 mice had metastasis of pelvic nodes at 12th day and 15th day respectively in the experimental group, and 4 and 4 respectively in control group. At the 15th day post operation, in control group, liver parenchyma metastasis and renal pelvis metastasis were observed in 1 and 1 case, respectively, while in experimental group, no distant metastasis was observed. The life span of tumor-bearing mice in the experimental and control groups was (15.80±0.84) and (16.00±0.71) days, respectively (P＞0. 05 ) . Conclusion Complete denervation of sympathetic adrenergic nerves seems to have no significant effects on the local growth, invasion and pelvic nodes metastasis of prostate cancer in C57BL/6 mice, but may have some inhibition effects on its distant metastasis.     

金属硫蛋白对皮瓣存活及细胞凋亡基因的影响

目的 观察金属硫蛋白(MT)对细胞凋亡基因及皮瓣存活影响.方法 大鼠背部形成随意皮瓣,实验组予以MT,对照组予以生理盐水,术后10 d测皮瓣存活率,术后3 d行苏木素-伊红(HE)染色光镜检查,激光多普勒测血运,术后3 d测皮瓣细胞凋亡基因bcl-2、bax蛋白.结果 MT组皮瓣的存活率(69.88±3.12)%高于对照组(60.65±2.98)%(P＜0.01).MT组中性粒细胞较多,激光多普勒血流量相对值(LDF值)下降少(P＜0.01).bcl-2蛋白表达MT组(2.98±0.23)较对照组(1.24±0.11)高(P＜0.01).bax蛋白(0.09±0.02)较对照组(0.23±0.09)低(P＜0.01).结论 皮瓣术后MT促进bcl-2,抑制bax,从而提高皮瓣存活率.

Abstract：

Objective To investigate whether metallothionein (MT) influences necrosis and expression of bcl-2 and bax in random pattern skin flaps of rats. Methods After flap operation, the rats in MT group were injected with MT, and those in control group received an injection of drug-free saline. Survival rate of the flaps was measured and the changes in the tissue were observed under the light microscopy.By using Doppler, the blood flow of flap was measured. The expression of bcl-2 and bax was detected. Results The survival area in MT group (69. 88 ±3. 12)% was significantly greater (P＜0.01) than that in control group (60. 65 ± 2. 98) %. There were more inflammatory cells in control group than in MT group.LDF in MT group was significiently higher than that in control group (P ＜0.01 ) 1st h, 2nd h, 3rd h, 6th h,1 st day, 3rd day, 6th and 7th day postoperation and at the 10th day postoperation (P ＜ 0. 05 ). At the 3rd day postoperation, the expression of bcl-2 in MT group (2.98 ± 0. 23 ) was significantly higher than that in control group ( 1.24 ± 0. 11 ) ( P ＜ 0. 01 ). The expression of bax in MT group ( 0. 09 ± 0. 02 ) was significantly lower than that in control group (0. 23 ± 0. 09) ( P ＜ 0. 01 ). Conclusion MT can affect the expression of bcl-2 and bax and significantly promote survival of skin flap.     

大鼠慢性皮肤溃疡创面愈合中转化生长因子-β1、胶原Ⅰ、胶原Ⅲ的蛋白表达

目的 观察大鼠慢性皮肤溃疡创面愈合过程中转化生长因子-β1( TGF-β1)、胶原Ⅰ和胶原Ⅲ的蛋白表达。方法 将24只8周龄雌性Wister大鼠分为单纯创面组(A组)和皮瓣+创面组即缺血模型组(B组),每组各12只;苏木素-伊红(HE)染色法观察创面1、3、7、10d上皮化率、收缩率及中性粒细胞;采用酶联免疫吸附试验(ELISA)方法测定创面1、3、7、10 d TGF-β1、Ⅰ型和Ⅲ型胶原的蛋白表达。结果 A组上皮化率在各个时间段均高于B组,且在第7天差异有统计学意义(P＜0.05)。A组收缩率明显低于B组。A组中性粒细胞第1、3天逐渐增加,第3天增加到最多,随后逐渐减少;B组在1、3、7d出现增加趋势,第7天增加到最多,第10天减少。TGF-31含量A组于术后1、3、7、10d呈曲线上升趋势,B组在术后1、3、7d逐渐减低,10 d较7d略有回升,且在第1天两组差异有统计学意义(P＜0.05)。胶原Ⅰ蛋白的含量两组随着术后时间的延长均呈减少趋势,在第10天两组差异有统计学意义(P＜0.05)。胶原Ⅲ蛋白的含量两组随术后时间的延长也呈减少的趋势,但在第3天A组比B组明显增加,差异有统计学意义(P＜0.05)。结论 在缺血的干预因素作用下TGF-β1、Ⅰ型和Ⅲ型胶原蛋白表达的减少可能延迟了慢性创伤的正常愈合。     

颅脑损伤后海马区notch信号分子及其相关miRNA的表达变化

目的 观察颅脑损伤后海马区notch1相关微小RNA(miRNA)、notch1及nestin的表达变化,探讨颅脑损伤后神经干细胞增殖机制.方法 采用自由落体法复制闭合性颅脑损伤小鼠模型,于伤后4 h、1 d和3 d处死动物.(1)实时聚合酶链反应(Real-time PCR)检测伤后4 h、1 d和3d伤侧海马区mir-326、mir-34a和notch1 mRNA的表达变化.(2)免疫荧光染色检测伤后3 d伤侧海马齿状回区notch1和nestin蛋白表达变化.结果 (1)颅脑损伤后4 h、1 d和3 d组海马区mir-326和mir-34a表达水平分别为假手术组的(0.36±0.16)、(0.16±0.04)、(0.36±0.17)倍和(0.48±0.15)、(0.50±0.04)、(0.25±0.14)倍,均低于假手术组(P＜0.01);(2)颅脑损伤后4 h、1 d和3 d组海马区notch1 mRNA表达水平分别为假手术组的(1.77±0.17)、(2.55±0.48)和(2.44±0.58)倍,均高于假手术组(P＜0.05);颅脑损伤后3 d组海马齿状回区notch1阳性细胞明显多于假手术组[(18.20±3.56)个比(0.40±0.55)个,P＜0.01].(3)颅脑损伤后3 d组海马齿状回区nestin阳性细胞数明显高于假手术组[(21.80±5.07)个比(0.80±0.45)个,P＜0.01].结论 在颅脑损伤后海马区神经干细胞增殖过程中,mir-326和mir-34a表达明显降低,其靶基因notch1 mRNA和蛋白表达明显升高,提示mir-326和mir-34a可靶向抑制notch信号分子,调控伤后神经干细胞增殖过程.

Abstract：

Objective To observe the expression of notch1-related miRNA, notch1 and nestin,and explore the regulatory mechanism of neural stem cells proliferation in hippocampus of mice after traumatic brain injury (TBI). Methods Mice were suffered closed head injury, and then sacrificed at 4th h,1st day and 3rd day post-injury. Real-time polymerase chain reaction (PCR) was used to detect the expression levels of mir-326, mir-34a and notch1 mRNA in mice hippocampus at 4th h, 1st day and 3rd day post TBI. Immunofluorescence was conducted to determine protein expression levels of notch1 and nestin in mice hippocampus at 3rd day post TBI. Results ( 1 ) Relative to sham group, the levels of mir-326 and mir-34a at 4th h, 1st day and 3rd day after TBI in the hippocampus were respectively (0. 36 ± 0. 16),(0. 16±0.04), (0.36±0. 17) and (0.48±0. 15), (0.50±0.04), (0.25 ±0. 14) fold (P＜0.01);(2) Relative to sham group, the levels of notch1 mRNA at 4th h, 1st day and 3rd day post TBI injury in the hippocampus were respectively (1.77 ±0. 17), (2.55 ±0.48) and (2.44±0.58) fold (P＜0.05).Notch1 + cells in the DG at 3rd day post TBI were significantly increased compared to sham group ( 18.20± 3.56 vs 0. 40 ±0. 55 ,P ＜0. 01 ); (3) Nestin + cells in the DG at 3rd day post TBI were increased apparently compared to sham group (21.80 ± 5. 07 vs 0.80 ± 0. 45, P ＜ 0. 01 ). Conclusion mir-326 and mir-34a may play a key role in trauma-induced neural stem cells proliferation in hippocampus in vivo by targeting notch1 signaling.     

不同来源的表皮干细胞促进皮肤创面愈合的研究

目的 观察表皮干细胞(ESCs)及去分化来源的表皮干细胞(DESCs)对皮肤创面愈合的促进作用及其异同.方法 由新鲜人包皮标本以贴壁法分离ESCs,随机分为3组,第1组作为原代ESCs:第2组传代培养至第6代时可见细胞成熟呈表皮细胞(ECs)形态,由原来的大克隆生长的小圆细胞形态变为较为肥大的不规则型细胞,无法形成克隆样增殖;其细胞表型也由β1整合素、CK19强阳性、CK10阴性变为β1整合素、CK19阴性,而CK10强阳性,予以100μg/L的碱性成纤维细胞生长因子(bFGF)培养48 h进行去分化诱导,48 h后ECs周边开始出现小圆细胞并呈克隆样生长,β1整合素、CK19、CK10等细胞表型也趋于与ESCs一致,即DESCs;第3组传代培养至第6代获得ECs.3组细胞皆以D-Hanks液重悬调其终质量浓度为2×106个/ml.将12只裸鼠每只背部左右各制备1个直径6 mm创面,共24个创面.将全部创面随机等分为4组,每组6个,分别以ESCs、DESCs、Ecs及生理盐水(NS)各0.2 ml进行创面注射.于术后0、3、7 d测量创面面积进行统计学分析,并于术后7 d行创面取材苏木素-伊红(HE)染色.结果 ESCs组与DESCs组在术后3 d时创面面积分别为(11.758±2.544)、(11.515±1.351)mm2,7 d时创面面积分别为(1.795±1.063)、(2.043±1.138)mm2,修复速度要明显快于ECs组与NS组[3 d时创面面积分别为(17.857±1.722)、(16.192±2.256)mm2,7 d时创面面积分别为(5.367±1.219)、(5.070±1.357)mm2,P＜0.01];而ESCs与DESCs组之间创面修复速度比较差异无统计学意义(P＞0.05);ECs组与NS组之间比较差异无统计学意义(P＞0.05).HE染色提示ESCs组与DESCs组的创面愈合质量优于ECs组与NS组,可见有皮肤附属腺的再生且纤维瘢痕组织较少.结论 ESCs与DESCs皆有促进创面愈合的作用,相互之间无明显差异.

Abstract：

Objective To investigate the enhancing effect of epithelial stem cells (ESCs) and dedifferentiation derived epithelial stem cells (DESCs) in the healing of epidermal wound. Methods ESCs were isolated from the fresh circumcised foreskins by using adherence method and randomly divided into three cohorts: ESCs cohort, DESCs cohort and epithelial cells (ECs) cohort. The final cell density was adjusted to 2 × 106/ml with D-Hanks in each group. The model of BALB/C mice full-thickness skin loss wounds was established. Two circular 6 mm-diameter skin loss wounds were cut on every BALB/C mouse back. Twenty-four wounds of 12 BALB/C mice were divided equally and randomly into 4 groups:DESCs, ESCs, ECs and NS. The cell suspensions of DESCs, ESCs and ECs were injected respectively into the wound edges with the density of 2 × 106/ml. And the volme was 0. 2 ml per treatment wound. The same volume of normal saline was injected in NS group. On the postoperative day 0, 3, 7, all of the wound areas were measured and analyzed statistically. On the postoperative 7 day, the sections were made and observed by using hematoxylin and eosin (HE) staining. Results The wound areas in DESCs group and ESCs group were ( 11. 758 ± 2. 544) and ( 11.515 ± 1.351 ) mm2 on the 3rd day postoperatively, and ( 1. 795 ±1. 063) and (2. 043 ± 1. 138) mm2 on the 7th day postoperatively, respectively. The wound areas in ECs group and NS group were ( 17. 857 ± 1. 722) and ( 16. 192 ±2. 256) mm2 on the 3rd day postoperatively,and ( 1. 795 ± 1. 063) and (2. 043 ± 1. 138) mm2 on the 7th day postoperatively, respectively. There was statistically significant difference in wound area between ESCs or DESCs group and ECs, NS groups on the 3rd, 7th day postoperatively ( P ＜ 0. 01 ), but there was no statistically significant difference between DESCs and ESCs groups ( P ＞ 0. 05 ). HE staining showed the repairing quality of treatment wounds was superior to that in the control wounds, and regenerating glands and less fibrous connective tissues were seen in ESCs and EDSCs groups. Conclusion Both ESCs and DESCs could promote the wound repair.     

高强度聚焦超声对淋巴管的生物学效应

目的 用高强度聚焦超声(HIFU)辐照大鼠淋巴管,观察产生的生物学变化,探讨HIFU治疗乳糜尿的可行性.方法 将SD大鼠32只随机均分为4组;用高强度聚焦超声辐照大鼠右侧腹股沟区,左侧对照;分别于辐照后24 h、72 h、7 d、30 d观察辐照区及周围组织肉眼及病理改变.结果 HIFU辐照后,大鼠腹股沟淋巴管24 h及72h组表现为管腔变形、破裂等急性损伤改变,7 d及30 d组表现为淋巴管数量减少、管腔闭塞、管周瘢痕形成;各时间组实验侧淋巴管密度(LVD)比自身对照侧明显减少:24 h组为1.100±0.428比1.450±0.411(P＜0.01);72 h组为0.725±0.238比1.575±0.362(P＜0.01);7 d组为0.375±0.198比1.575±0.249(P＜0.01);30 d组为0.175±0.198比1.500±0.400(P＜0.01);辐照后,随时间延长,实验侧LVD明显减少(F=16.669,P＜0.01);辐照后7 d,LVD减少不明显(P＞0.05).结论 高强度聚焦超声可靶向损伤淋巴管,最终使管腔粘连闭塞,因此高强度聚焦超声治疗乳糜尿有可行性.

Abstract：

Objective To investigate the use of high-intensity focused ultrasound ( HIFU) to noninvasively produce biological effect in rat lymphatic vessels in vivo, and the feasibility of HIFU in the treatment of chyluria. Methods Thirty-two SD rats were divided into four groups randomly. The right inguinal regions were irradiated and the left served as controls. At the 1st, 3rd, 7th and 30 day after HIFU irradiation, the gross and pathological changes were observed. Results There were acute changes such as the lymphatic vessels rupture and deformation on the experimental sides at 1st and 3rd day after HIFU irradiation. The number of lymphatic vessels was reduced, lumen was occluded, scar formed around the lumen at 7th and 30th day. The lymphatic vessec density (LVD) on the experimental side was significantly decreased as compared with the controls after HIFU irradiation (experimental sides vs controls: 1st day,1.100 ±0.428 vs 1.450 ±0.411 (P＜0.01); 3rd day, 0. 725 ±0. 238 vs 1.575 ±0.362 (P＜0.01);7th day, 0. 375 ± 0. 198 vs 1. 575 ± 0. 249 ( P ＜ 0. 01); 30th day, 0.175 ± 0.198 vs 1. 500 ± 0.400 (P＜0. 01). There was a significant decrease in LVD on the experimental side with the time going (F = 16.669,P＜0.01). However, there was no significant decrease in LVD at the 7th day (P＞0.05).Conclusion HIFU could precisely damage lymphatic vessels of rats and make lumen occlusion finally,suggesting the feasibility of HIFU for chyluria treatment.     

雌激素对大鼠创面愈合过程中微血管形成的影响及其机制

目的 观察雌激素对大鼠全层皮肤缺损创面愈合过程中微小血管再形成的影响.方法 将40只雌性Wistar大鼠随机分成3组:单纯损伤组15只(A组),去卵巢组15只(B组)和假手术组10只(C组).8周后,在背部制成全层全损皮肤创伤愈合模型,分别于伤后1、3、7、14和21 d记录3组各个时间点伤腔容积,并在创缘取材.常规组织学观察皮肤愈合过程和微血管的数量,同时采用雌激素β受体(ER-β)进行免疫组织化学染色,观察微血管及周围细胞受体的染色.结果　比较各个时间点的伤腔容积,单纯损伤组和假手术组大鼠创面内肉芽组织形成量明显大于去卵巢组,组织学评分略高于去卵巢大鼠;而单纯损伤组和假手术组之间差异无统计学意义(P＞0.05).卵巢切除大鼠伤后各时间点,真皮内微小血管的数量较单纯损伤组减少,各组均于伤后7 d,随着成纤维细胞和毛细血管数目的 增加,肉芽组织生长达到高峰.同时,各损伤组创基内微小血管周围均出现较为幼稚的细胞,其表面可以表达ER-β,但卵巢切除组(159.91±12.65)稍弱于单纯损伤组(189.36±27.32),差异无统计学意义(P＞0.05).结论　大鼠去卵巢后创面愈合速度明显减慢与微血管形成有关,雌激素及其受体在该过程中有重要作用.

Abstract：

Objective To investigate the effect of estrogen on the revascularization of full-thickness skin wounds healing in rats. Methods Forty 2-month-old female Wistar rats were divided into three groups: control group (group A, pure injury,n = 15), ovariectomized group (group B,n = 15) and sham operation group (group C,n = 10 ). After 8 weeks, full-thickness wounds model, 18 mm in diameter, were made on the back of the each animal. The volume of wound was measured at each time point. The specimens of wound edge were obtained at the 1st, 3rd, 7th, 14th and 21st day after injury. The process of wound healing and the number of microvessels in the granulation tissue were evaluated by Haematine-Eosin (HE) staining. The immunohistochemical staining was used to detect the expression of estrogen receptor β (ER-β) in microvascular endothelial cells and their surrounding cells. Results To compare the volume of wound at each time point, the granulation tissue formation in group A and group C was significantly higher than that in group B, but there was no significant difference between group A and group C. The number of capillary blood vessels in group B was significantly reduced as compared with that in group A. The expression of ER-β was positive on the cells surface in all groups. ER-β expression in group B was weaker than in group A ( 159. 91 ± 12. 65 v. s 189. 36 ±27. 32,P ＜0. 05). Conclusion The delay of wound healing in ovariectomized rats has a great relationship with angiogenesis. Estrogen and its receptor might play important roles in the process.     

CTLA4-Ig对肝细胞移植大鼠免疫耐受的作用及其机制

目的 探讨细胞毒性淋巴细胞相关抗原4融合蛋白(CTLA4-Ig)诱导肝细胞移植大鼠免疫耐受的作用及机制.方法 10%D-氨基半乳糖(D-gal)一次性腹腔注射建立大鼠急性药物性肝衰竭模型;采用肝脏原位灌注法分离纯化肝细胞,经脾脏移植后随机分为两组.实验组腹腔一次性注射CTLA4-Ig,对照组不予处理.两组均分别于术后第1、3、5、7天采外周血观察白细胞介素(IL)-2、肿瘤坏死因子(TNF)及肝功能变化;术后1周测两组大鼠外周血T细胞亚群,处死大鼠后取脾脏苏木素-伊红(HE)染色.结果 实验组谷丙转氨酶(ALT)、血清总胆红素(TBil)于术后第7天分别为(6.5±7.3)IU/ml、(5.1±1.6)mmol/L,低于对照组.术后治疗组IL-2含量明显下降,第7天达到(1. 3138±0.8508)ng/L,两组差异有统计学意义(P＜0.05);术后TNF含量两组之间差异无统计学差异(P＞0.05).外周血CD4+T细胞、CD4+/CD8+T细胞实验组分别为(37.3±7.2)%、(1.5±0.1)%,低于对照组(P＜0.05),CD8+T细胞两组差异无统计学意义(P＞0.05).术后第7天治疗组脾内仍可见肝细胞或肝细胞团,对照组见大量淋巴细胞浸润,但很少见肝细胞.结论 CTLA4-Ig能诱导经脾同种异体肝细胞移植大鼠免疫耐受,使急性肝衰大鼠肝功能得到改善.可能是抑制T淋巴细胞亚群,且主要是抑制CD4+T细胞,使CD4+/CD8+T细胞比值下降;抑制IL-2的分泌.

Abstract：

Objective To investigate the immunosuppressive effect of cytotoxic T Iymphocyte associated antigen 4 Ig fusion protein (CTLA4-Ig) in rat allograft hepatocyte transplantation model and the mechanisms. Methods Acute liver failure (ALF) model was established by intraperitoneal injection of 10% D-gal solution to SD rates. Collagenase perfusion was performed on SD rats to separate liver cells. SD rats with ALF were subjected to intrasplenic hepatocyte transplantation and randomly divided into two groups. The experimental group received intraperitoneal injection of CTLA4-Ig. The concentrations of interleukin (IL)-2 and tumor necrosis factor (TNF), liver function and histologicy were observed at the 1st,3rd, 5th, 7th day after operation and the T lymphocyte subsets were detected by using immunohistochemistry at the 7th day after operation. Results The levels of ALT and TBil were respectively (6. 5 ±7.3) IU/ml and (5.1 ± 1.6) mmol/L at the 7th day after operation and significantly decreased after injection of CTLA4-Ig ( P ＜ 0. 01 ). IL-2 concentration in the experimental group was ( 1.3138 ± 0. 8508 ) ng/L at the 7th day after operation and significantly decreased (P ＜0. 05). TNF had no significant difference between two groups after operation ( P ＞ 0. 05 ). T lymphocyte subsets, mainly CD4 + , in the experimental group was significantly decreased as compared with control group ( P ＜ 0. 05 ), so did the CD4 +/CD8 +. Histological changes: At the 7th day after operation, there were some hepatocytes in the spleen of the experimental group. But in the control group, the changes in the spleen were characterized by severe lymphocyte infiltration. There were no hepatocytes both groups. Conclusion CTLA4-Ig can induce rat allogeneic hepatocytes intrasplenic transplantation immune tolerance. It may improve the liver function of rats with ALF.CTLA4-Ig can decrease T lymphocyte subsets, mainly CD4 + and concentrations of IL-2.     

膨体聚四氟乙烯覆盖创面的实验研究

目的 了解高分子材料膨体聚四氟乙烯(ePTFE)作为创面覆盖材料的可行性.方法 将45只SD大鼠背部制成全层皮肤至筋膜层损伤创面后分为:自体移植组,创面行自体皮片回植;异体移植组,以15只Wistar大鼠为供皮鼠,将皮片移植于SD大鼠创面;实验组,在创面上覆盖ePTFE.每组15只SD大鼠.肉眼观察各组大鼠创面愈合情况.于术后3、7、14 d切取各组大鼠创面组织标本行HE染色,光学显微镜下观察组织切片中巨噬细胞、Fb、淋巴细胞数量.免疫组织化学法检测γ干扰素(IFN-γ)和IL-2表达水平.结果 自体移植组、实验组大鼠创面愈合良好,未见红肿及感染征象;异体移植组大鼠术后8 d出现排斥反应,表皮有不同程度的变性坏死,创缘红肿.各时相点异体移植组镜下可见巨噬细胞、Fb、淋巴细胞数量均高于实验组和自体移植组(P＜0.01).免疫组织化学染色可见,术后7 d异体移植组IFN-γ和IL-2表达的平均灰度值分别为129±7、113.7±2.7,均明显低于自体移植组(189±6、180.3±3.7,P＜0.01)和实验组(144±8、137.3±1.9,P＜0.01).结论 ePTFE可引起较小的炎性反应及异物反应,置于受损创面上未见不良反应,可以将其作为创面覆盖材料.     

明胶-白芨胶/丹参材料促进大鼠损伤组织内血管内皮生长因子及转化生长因子-β1表达的效果

目的 观察明胶-白芨胶/丹参材料内丹参浓度对损伤组织血管内皮生因子(VEGF)及转化生长因子(TGF)-β1表达的促进效果.方法 制备明胶-白芨胶/丹参多孔材料.175只大鼠随机分为5组,每组35只.4个实验组大鼠皮下依次植入含丹参浓度为1、2、4、5 ml/100 g的材料,对照组植入单纯明胶-白芨胶多孔材料.术后第1、3、7、14、21、28和56天,每组随机选5只大鼠处死.将含周围组织的材料取出,苏木素-伊红(HE)染色切片.将术后1、3、7、14 d的标本行VEGF及TGF-β1抗体免疫组织化学染色,利用Image J软件,镜下观察分析.结果 材料约8周完全降解,HE染色未示异常反应.免疫组织化学染色阳性点计数显示,实验组VEGF及TGF-β1表达明显强于对照组(P＜0.01).在表达最强的第3天,丹参浓度为2 ml/100 g时,表达值分别为457.7±12.7和1099.7±22.4.丹参浓度超过2 ml/100 g时,实验组间VEGF及TGF-β1表达差异无统计学意义(P＞0.05).结论 明胶-白芨胶/丹参多孔材料具有良好的组织相容性,丹参在材料内促进VEGF及TGF-β1表达的适宜浓度约为2 ml/100 g.