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.     

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.     

K+、Cl-离子通道阻滞对体外培养兔关节软骨细胞基质合成代谢的影响

目的 观察阻断体外培养兔关节软骨细胞膜上的K+、Cl-离子通道对其糖胺多糖(GAG)、Ⅱ型胶原基质合成代谢的影响.方法 2个月龄新西兰白兔,无菌条件下切取双膝关节软骨,酶解法分离软骨细胞,以5 ×104/孔接种于24孔板中.正常培养2 d细胞贴壁后换液,并以12个孔为1组随机分为3组,对照组:用DMEM/F12正常培养;K+离子通道阻滞组(简称4-AP组):利用含1 mmol/L 4-氨基吡啶(4-AP)的DMEM/F12培养,特异性阻断电压门控型K+离子通道;Cl-离子通道阻滞组(简称SITS组):利用含1 mmol/L4-乙酰氨基-4'-异硫氰基-2,2'-乙拌磷酸均二苯乙烯(SITS)的DMEM/F12培养,阻断阴离子通道,主要是Cl-离子通道.分别在换液后第3、6、9天留取各孔上清液并分为2份,1份以阿尔新蓝法检测其GAG的含量,同时另1份以酶联免疫吸附试验(ELISA)法检测Ⅱ型胶原的含量(n=12).结果 4-AP组在第3天时与对照组比较GAG含量明显下降(P＜0.05),但第6天和第9天差异无统计学意义(P＞0.05),同时在第3天和第6天Ⅱ型胶原含量显著增加(P＜0.05),第9天时有下降趋势,但差异无统计学意义(4-AP组GAG含量分别为17.23±1.47、20.98±2.61、33.13±8.13;Ⅱ型胶原含量分别为0.793±0.214、0.789±0.084、0.715±0.388);SITS组在第3、6、9天与对照组比较GAG含量都显著增加(P＜0.05),同时在第3天和6天Ⅱ型胶原含量显著增加(P＜0.05),第9天仍然有增加的趋势,但差异无统计学意义(SITS组GAG含量分别为54.30±4.43、56.47±3.14、58.71±2.50;Ⅱ型胶原含量分别为0.793±0.125、0.853±0.060、0.925±0.053).结论　阻断K+、Cl-离子通道后显著促进软骨细胞GAG、Ⅱ型胶原的合成,尤其是阻断Cl-离子通道后,GAG的合成增加尤为明显.

Abstract：

Objective To observe the effects of potassium and chloride channel blockers on the glycosaminoglycan (GAG) and collagen type Ⅱ synthesis of rabbit articular chondrocytes in vitro. Methods Two-month-old New Zealand rabbits were killed and their knee joints were taken out under the aseptic condition. The chondrocytes were isolated by enzymolysis method and cultivated into a 24-well plate ( seeded 5 × 104 cells per well). Then the chondrocytes were divided into three groups randomly after culture for 2 days when the cells were adherent. In the control group the cells were cultured by DMEM/F12, in the potassium channel blocking group [4-aminopyridine (4-AP) group] the cells were cultured in the medium containing 1 mmol/L 4-AP, and in the chloride channel blocking group [4-acetamido-4-isothiocyano-2,2-disulfonic acid stilbene (SITS) group] the cells were cultured in the medium containing 1 mmol/L SITS.The GAG synthesis was measured by Alician Blue method and collagen type Ⅱ synthesis was estimated by the enzyme linked immunosorbent assay (ELISA) in the media at the 3rd, 6th and 9th day after medium change ( n = 12). Results As compared with the control group, the GAG content in 4-AP group was significantly decreased at the 3rd day ( P ＜ 0. 05 ), but showed no significant difference at 6th and 9th day (P ＞0. 05); the collagen type Ⅱ content was significantly increased at 3rd and 6th day (P ＜0. 05), and showed a decrease tendency at 9th day without significant difference ( GAG: 17.23 ± 1.47, 20. 98 ± 2. 61,33. 13 ± 8. 13; collagen type Ⅱ: 0. 793 ± 0. 214, 0. 789 ± 0. 084, 0. 715 ± 0. 388). The GAG content in the SITS group was significantly increased at the 3rd, 6th and 9th day (P ＜0. 05), while the collagen type Ⅱ content was significantly increased at 3rd and 6th day ( P ＜0. 05), and showed an increase tendency at 9th day without significant difference ( GAG: 54. 30 ±4. 43, 56. 47 ± 3. 14, 58.71± 2. 50; collagen type Ⅱ: 0. 793 ±0. 125, 0. 853 ±0. 060, 0. 925 ±0. 053). Conclusion The blockage of potassium and chloride channels can increase the GAG and collagen type Ⅱ synthesis of chondrocytes in vitro, especially when chloride channel is blocked, the GAG synthesis was significantly increased.     

联合内皮祖细胞移植对小鼠骨髓移植预处理中肝脏内皮损伤的修复作用

目的 研究联合内皮祖细胞(EPC)移植对小鼠骨髓移植预处理中肝脏内皮损伤的修复作用.方法 将C57BL/6小鼠分为4组,每组10只.(1)正常对照组:小鼠不做任何处理,仅作为正常对照;(2)单纯照射组:单次给予全身照射(TBI)预处理,不进行骨髓移植;(3)单纯移植组:给予单纯照射组相同的TBI预处理,TBI后4 h内经小鼠尾静脉输注C57BL/6小鼠骨髓单个核细胞5×106/只;(4)联合移植组,小鼠的处理方式与单纯移植组相同,仅在骨髓移植的同时经尾静脉输注C57BL/6小鼠EPC 5×105/只.TBI后第2、4、7、14、21天,检测各组小鼠肝脏重量的变化情况,并于TBI后第4、7、14、21天对各组小鼠肝脏进行组织病理学检查.结果 单纯照射组、单纯移植组和联合移植组小鼠肝脏重量均于TBI后第2天开始明显增加,于第14天达到高峰,峰值分别为正常对照组的(1.65±0.15)倍(P＜0.05)、(1.61±0.06)倍(P＜0.05)和(1.11±0.4(0)倍(P＜0.05);以后均呈下降趋势,第21天时单纯照射组和单纯移植组肝脏重量仍明显高于正常对照组(P＜0.05),但联合移植组小鼠肝脏重量已完全恢复正常.组织病理学检查显示,单纯照射组小鼠肝窦内皮损伤明显,肝细胞水肿及严重的炎症细胞浸润,第7天时肝细胞水肿、坏死较前明显加重,几乎无存活的肝血窦内皮细胞;第14天时单纯移植组小鼠肝窦内皮损伤较前有所减轻,但到第21天时仍未恢复正常;联合移植组小鼠第7天时肝窦内皮及肝细胞水肿、坏死程度均较轻,到第14天时已基本恢复正常.结论 造血干细胞移植前的预处理会造成受者肝脏内皮损伤,且此损伤持续存在;移植时联合输注EPC能修复肝窦内皮的损伤.

Abstract：

Objective To study the repair function of united endothelial progenitor cells (EPC)transplantation on injured liver endothelium by bone marrow transplantation (BMT) conditioning.Methods C57BL/6 mice were divided into four groups randomly: normal control group, without any treatment; irradiation alone group, administered a total body irradiation(TBI) pretreatment, without BMT; (3) BMT alone group: C57BL/6 mice were infused with bone marrow mononuclearcells (MNC) 5 × 106/only through caudal vein not more than 4 h after the same TBI pretreatment as the irradiation alone group; united transplantation group: receiving the same way as the BMT alone group, but C57BL/6 mice were infused with EPC 5 × 105/only at the same time. Two, 4, 7, 14, and 21 days after the TBI, the changes of the liver weight were observed regularly. The histopathological examination of liver was done at the 4th, 7th, 14th, and 21st day after the TBI. Results In irradiation alone group, BMT alone group and united transplantation group the liver weight began to increase significantly on the day 2 and peaked at 14th day after the TBI, and the peaks were respectively (1.65±0. 15) times (P＜0. 05), (1.61 ±0.06) times (P＜0.05), and (1.11 ±0.40)times (P＜0. 05) of those in normal control group. At the day 14, the liver weight in irradiation alone group, BMT alone group and united transplantation group began to decrease, and on the day 21 the liver weight in united transplantation group had been completely restored to normal level, however the liver weight in irradiation alone group and BMT alone group were still significantly heavier than that in normal control group (P＜0. 05). Liver histopathological examination revealed that there were obvious sinusoidal endothelial cells (SEC) injury, hepatocyte edema and severe inflammatory cell infiltration in irradiation alone group, and on the day 7 the hepatocyte edema and necrosis were significantly worse than before, and almost no alive SEC were found. On the day 14 the injury of SEC in BMT alone group was lighter than before, but on the day 21 the injury had not returned to normal. On the day 7 the injury of SEC, hepatocyte edema and necrosis were alleviated in united transplantation group as compared with irradiation alone group and BMT alone group, and on the day 14 the injury had returned to normal basically. Conclusion The transplantation conditioning could damage recipient liver endothelium and the injury would persist, and united EPC infusion could repair the injured SEC following BMT.     

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.     

脑创伤患者急性期皮质醇分泌的变化

目的 探讨脑创伤患者急性期皮质醇(COR)分泌的变化.方法 创伤后2～24 h入院的脑创伤患者75例,根据Glasgow昏迷评分分为轻度脑创伤组(TBI1组,n=30)、中度脑创伤组(TBI2组,n=12)和重度脑创伤组(TBI3组,n=33),13例同期住院的颈椎病或颅骨骨瘤患者为对照组(C组).于入院后1 d时采集静脉血样,测定血清总COR、促肾上腺皮质激素和皮质醇结合球蛋白的浓度,计算游离COR浓度及游离COR指数.记录高血COR的发生情况.结果 与C组比较,TBI1组、TBI2组和TBI3组血清总COR、促肾上腺皮质激素、游离COR的浓度及游离COR指数均升高(P＜0.05),且TBI2组和TBI3组高于TBI1组(P＜0.05);四组间皮质醇结合球蛋白浓度比较差异无统计学意义(P＞0.05).TBI1组、TBI2组和TBI3组高血COR发生率高于C组,且TBI3组高于TBI1组和TBI2组(P＜0.05).结论 脑创伤患者急性期COR分泌升高,COR分泌水平与创伤程度有关.

Abstract：

Objective To investigate the changes in cortisol (COR) secretion in the acute phase of traumatic brain injury (TBI) .Method Seventy-five patients admitted to the hospital at 2-24 h after TBI were divided into 3 groups based on the Glasgow Coma Scale score: mild TBI group (group TBI1, n = 30), moderate TBI group (group TBI2, n = 12) and severe TBI group (TBI3, n = 33). Thirteen patients with cervical spondylosis or osteoma of the skull (admitted to the hospital at the same period) were regarded as control group (group C). Venous blood samples were taken on the first day after admission to measure the serum concentrations of total COR, adrenocorticotropin (ACTH) and corticosteroid-binding globulin (CBG). Free COR concentrations and free COR index were calculated. High blood COR was recorded. Result Compared with group C, the serum concentrations of total COR and ACTH, free COR levels and free COR index were significantly increased in TBI1, TBI2 and TBI3groups (P ＜ 0.05). The parameters mentioned above were significantly higher in TBI2 and TBI3 groups than in TBI1 group ( P ＜0.05). There was no significant difference in serum CBG concentrations among the four groups.The incidence of high blood COR was significantly higher in TBI1, TBI2 and TBI3 groups than in C group, and in TBI3 group thanin TBI1 and TBI2 groups (P ＜0.05). Conclusion COR secretion is increased in the acute phase of TBI and the level of COR secretion is related to the severity of brain damage.     

促红细胞生成素动员骨髓间充质干细胞治疗脊髓损伤

目的 观察促红细胞生成素(EPO)动员骨髓间充质干细胞(BMSCs)对脊髓损伤局部起的修复作用,并探讨其机制.方法 将30只SD大鼠随机分成假手术组、生理盐水组及EPO治疗组,各组均经尾静脉注射Hoechst33342标记的BMSCs,移植后1、3、7、14、21、28 d采用BBB法进行大鼠后肢运动功能评分观察大鼠神经功能恢复情况,应用免疫荧光检测比较各组BMSCs动员到脊髓损伤处的数目,Western blot检测损伤局部BMSCs表达脑源性神经营养因子(BDNF)和神经生长因子(NGF)的水平.结果　从第3天起,EPO治疗组BBB评分分别为(5.29±0.69)、(8.18±0.38)、(13.32±0.77)、(15.25±1.83)、(18.71±1.54),明显高于生理盐水组,移植的BMSCs计数分别为(90.12±7.68)、(116.26±13.54)、(186.32±20.35)、(211.64±31.83)、(298.71±21.54),数目明显增加(P＜0.05),其BDNF和NGF的表达也明显增强(P＜0.05).结论 EPO能够动员BMSCs向脊髓损伤部位迁移并通过促进BMSCs表达BDNF和NGF来促进神经功能的恢复.

Abstract：

Objective To obsevre the effect of erythropoietin (EPO) treatment on the mobility of bone marrow mesenchymal stem cell (BMSCs) to the sites of injured spinal cord and explore the molecular mechanism. Methods Thirty SD rats were randomly divided into sham-operation group, saline group and EPO treatment group. BMSCs labeled with Hoechst33342 were transplanted by tail vein. Functional outcome measurements were evaluated by the Basso, Beattie and Bresnahan (BBB) score at 1st, 3rd, 7th,14th, 21st and 28th day. The immigrating number of BMSCs was measured by immunofluorescent staining.The expression of brain-derived neurotrophic factor (BDNF) and nerve growth factor (NGF) in BMSCs was detected by Western blotting. Results From the 3rd day on, the BBB scores in EPO treatment group were (5.29±0.69), (8.18±0.38), (13.32±0.77), (15.25 ±1.83), (18.71 ±1.54) respectively, which were obviously higher than in saline group. The number of BMSCs was (90. 12 ±7. 68), (116. 26 ± 13. 54),( 186. 32 ± 20. 35 ), (211.64 ± 31.83 ) and (298.71 ± 21.54) respectively, which was significantly increased in EPO treatment group (P ＜ 0. 05 ). The protein levels of BDNF and NGF expressed by BMSCs were increased significantly (P ＜ 0. 05) in EPO treatment group as compared with sham-operation group.Conclusion EPO treatment after spinal cord injury may help BMSCs migrate to the injury site and accelerate recovery of neural function by up-regulating the expression of BDNF and NGF.     

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

目的 观察荷负电气溶胶治疗对大鼠烫伤创面愈合过程中白细胞介素(IL)-8、IL-10表达的影响,探讨荷负电气溶胶治疗促进创面愈合的作用机制.方法 制作SD大鼠深Ⅱ度烫伤模型,采用分组对照方法,将40只鼠随机分为治疗组(n1=20)和对照组(n2=20).治疗组应用荷负电气溶胶治疗,每次1.5 h,每天2次,直至创面愈合;对照组不作荷负电气溶胶治疗.伤后第1～11天分别取创面标本制作切片,采用免疫组织化学和图像分析方法,检测创面愈合过程中IL-8和IL-10表达水平.结果 创面平均愈合时间治疗组为(7.00±1.15)d,对照组为(9.00±1.34)d,治疗组创面愈合时间明显提前(P＜0.01).免疫组织化学显示,两组IL-8均在伤后第1天开始表达.主要位于多核粒细胞和单核细胞;第3天表达明显增多达高峰,并见大量成纤维细胞表达,治疗组的峰值明显低于对照组,差异有统计学意义(P＜0.01),第5～11天表达水平迅速下降.两组IL-10伤后第1天在淋巴细胞和单核细胞均有表达;第3天开始有角质细胞表达并达高峰,第5～11天表达水平缓慢下降,但治疗组要明显高于对照组,第3～11天差异有统计学意义(P＜0.01).结论 荷负电气溶胶治疗能有效抑制创面IL-8的表达及促进IL-10的表达,缩短炎症进程,从而加速创面愈合.

Abstract：

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.     

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

目的 观察完全去交感肾上腺素能神经支配对C57BL/6小鼠前列腺癌生物学行为的影响.方法 雄性10周龄C57BL/6小鼠50只,平均分为2组.小鼠前列腺双侧背侧叶包膜下注入RM-1细胞0.5×106个,然后切断(实验组)或不切断(对照组)其双侧下腹下神经,术后6 d始,动态观察小鼠前列腺癌局部生长、盆腔淋巴结转移、远处转移和荷瘤生存期;切断的神经纤维行酪氨酸羟化酶免疫组织化学染色.结果 切断的神经纤维组织切片酪氨酸羟化酶免疫组织化学染色为强阳性.术后小鼠前列腺肿瘤形成并进行性增大,实验组与对照组小鼠前列腺体积术后6 d[(0.034±0.008)/(0.034±0.008)cm3]、9 d[(0.339±0.040)/(0.316±0.050)cm3]、12 d[(0.829±0.090)/(0.824±0.071)cm3]、15 d[(1.169±0.093)/(1.236±0.103)cm3],差异均无统计学意义(P＞0.05);术后12 d:实验组4例、对照组3例肿瘤侵犯盆壁肌肉,实验组3例、对照组4例出现盆腔淋巴结转移;术后15 d:两组均有4例肿瘤侵犯精囊、膀胱或发生盆腔淋巴结转移,对照组出现肝实质和肾盂转移各1例,实验组无远处转移病例发现.实验组和对照组小鼠荷瘤生存期分别为(15.80±0.84)d和(16.00±0.71)d,差异无统计学意义(P＞0.05).结论 完全去交感肾上腺素能神经支配对C57BL/6小鼠前列腺癌局部生长、侵袭和盆腔淋巴结转移等过程似无明显影响,对其远处转移可能有抑制作用.

Abstract：

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.     

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。     

慢性肾功能损伤的肝移植受者转换为西罗莫司治疗的疗效观察

目的 观察慢性肾功能损伤的肝移植受者转换为西罗莫司治疗的疗效.方法 应用钙调磷酸酶抑制剂(CNI)并伴有慢性肾功能损伤的肝移植受者23例(其中19例应用他克莫司,4例应用环孢素A)转换为西罗莫司(SRL)治疗.SRL的起始剂量为4mg/d,次日为2 mg/d,应用高压液相色谱法测定全血SRL浓度,当血SRL浓度达5～8 μg/L后,停用CNI类药物,同时服用吗替麦考酚酯,1 g/d.记录受者入组前的基础血清肌酐(Cr)、肌酐清除率、肾小球滤过率(GFR),并分别于用药后第1、3、6、12和24个月时监测血SRL浓度、Cr、肌酐清除率、GFR,同时监测受者体重、血压、血细胞计数、肝功能和肝脏生化指标、血脂、尿蛋白.于用药后12个月时行肝脏穿刺活检确认有无排斥反应.结果 23例平均随访29.4个月,随访期内死亡2例,另21例于用药后1、3、6、12和24月时的Cr分别为(147.40±23.36)、(152.60±20.08)、(150.20±22.64)、(137.60±18.09)、(138.30±17.04)μmol/L,与Cr的基础值[(158.91±29.13)μmol/L]相比较,1、12、24个月时的差异有统计学意义(P＜0.05).用药后1、3、6、12和24月时的肌酐清除率分别为(0.97±0.18)、(0.99±0.14)、(1.00±0.17)、(1.07±0.29)、(1.14±0.12)ml/s,与基础肌酐清除率[(0.91±0.14)ml/s]相比较,1、12、24个月时的差异有统计学意义(P＜0.05).用药后1、3、6、12和24月时的GFR分别为(0.80±0.15)、(0.78±0.11)、(0.75±0.12)、(0.84±0.10)、(0.94±0.13),与基础GFR[(0.71±0.11)ml/s]相比较,1、12、24个月时的差异有统计学意义(P＜0.05).应用SRL后第1、3、6、12和24个月时,Cr≤123μmol/L者所占的比例分别为38.1％、33.3％、28.6％、47.6％和52.4％.随访期内无受者发生排斥反应.结论 慢性肾功能损伤的肝移植受者转换为西罗莫司治疗可改善其肾功能.转换治疗未增加排斥反应的发生率.     

脊柱内窥镜下治疗极外侧型腰椎间盘突出症的临床疗效探讨

目的：探讨侧路脊柱内窥镜下治疗极外侧型腰椎间盘突出症的临床疗效。方法采用经皮脊柱内窥镜侧路手术治疗极外侧型椎间盘突出症患者32例,分别于术后1 d、1周、3个月、6个月采用疼痛视觉模拟量表（visual analogue scale, VAS）评分和Oswestry功能障碍指数（Oswestry disability index, ODI）评价疼痛及功能障碍情况,末次随访时采用Macnab标准评价临床疗效。结果术中未出现血管神经损伤等并发症,单节手术时间为20 min（10~35 min）,住院时间为5 d（3~7 d）。随访1 d、1周、3个月、6个月时的VAS评分分别为（2.53±0.61）分、（1.50±0.43）分、（1.48±0.35）分、（1.47±0.31）分,与术前的（8.81±0.42）分相比,差异均有统计学意义（均P＜0.05）;随访1 d、1周、3个月、6个月时的ODI分别为（18.01±6.42）%、（17.54±5.53）%、（16.33±6.42）%、（16.04±6.51）%,与术前的（76.51±13.25）%相比,差异均有统计学意义（均P＜0.05）。Macnab疗效评定结果,优19例,良11例,可2例,优良率为93.75%。结论经皮侧路脊柱内窥镜治疗极外侧型腰椎间盘突出症具有手术时间短、创伤小、恢复快等优点,疗效确切。     

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.