自体移植脾组织VEGF、KDR表达与血管再生的实验研究

目的 研究自体移植脾组织血管再生及VEGF、KDR表达规律，阐明VEGF、KDR对移植脾组织血管再生的调控作用，为脾脏外科临床及实验研究提供理论依据。方法 健康Wistar大鼠70只，体重100—120g，随机分为7组，每组10只中又设脾切除自体脾移植组5只，假手术组5只，分别于术后7，14，30，60，90，120，180d进行：(1)自体移植脾组织病理学检测；(2)大鼠行主动脉插管灌注墨汁，光镜观测再生血管并采用图像分析测定其密度；(3)免疫组化抗VEGF、KDR抗体染色，图像分析定量，阐明其表达规律及与血管再生的关系。结果 (1)自体脾组织移植术后7d即有血管从大网膜向脾组织内伸展，移植脾组织内血管密度逐渐增大，至术后180d血管再生接近正常；(2)自体脾组织移植术后7d、14d，VEGF、KDR阳性染色细胞密度迅速升高，术后60d达高峰，以后逐渐降低，至术后180d VEGF、KDR阳性染色细胞密度趋向正常。结论 自体脾组织大网膜内移植术是简便有效的脾移植方法；移植脾组织新生血管由大网膜再生而来；术后移植脾组织内VEGF、KDR表达量升高，促进血管形成，血管再生完成后恢复正常水平。     

自体移植脾组织神经肽Y阳性神经纤维再生的实验研究

目的：研究自休脾组织移植术后不同时相神经肽Y阳性（NPY^ ）神经纤维的再生规律。方法：健康Wistar大鼠56只，雌雄不限，体重100-120g，随机分为实验组及假手术组，术后7，14，30，60，90，120，180d取两组脾组织，行免疫组化抗NPY抗体染色及图像分析定量测定。结果：术后30d内，自体移植脾组织无NPY^ 神经纤维；术后60d，NPY^ 神经纤维出现于移植脾组织周边区域；术后90d，NPY^ 神经纤维向移植脾组织中央实质区伸展；术后120-180d，移植脾组织内NPY^ 神经纤维分布及密切均接近正常。结论：自体脾组织移植后可实现NPY^ 神经纤维再生；再生神经纤维来源于移植脾组织周围大网膜。     

腹膜后自体脾移植治疗外伤性脾破裂的前瞻性研究

目的 探讨腹膜后自体脾移植在严重脾外伤保脾手术中应用的可行性及价值。方法 采用前瞻性病例对照研究,将66例严重脾外伤病人分为腹膜后自体脾移植组例、大网膜自体脾移植组,其中腹膜后自体脾移植组32例,大网膜自体脾移植组34例。观察术后一般情况,术后1天～ 12个月血常规,血IgA、IgM、IgG、C3、Tuftsin水平变化。结果 腹膜后自体脾移植组手术时间短于大网膜自体脾移植组（P<0.05）,术后其它一般情况变化、术后1天～ 12个月血IgA、IgM、IgG、C3、Tuftsin水平两组无统计学差异（P> 0.05）。结论 腹膜后自体带蒂脾移植术能够保留脾脏的部分免疫功能,且手术操作简便,在临床上推广应用是可行的。     

隔离小肠段,网膜内自体脾组织移植的研究

目的:观察隔离小肠段、网膜内自体脾移植的抗感染能力并比较移植脾与原脾差异.方法:将24只雄性大耳白兔分为四组:Ⅰ、假手术组;Ⅱ、空肠内移植组;Ⅲ、网膜内移植组;Ⅳ、脾切除组.5个月后用1.4×10~(12)CFU/LⅢ型肺炎球菌静脉攻击.光镜、电镜观察,IBAS图象分析仪测红髓、白髓、边缘带面积.结果:移植脾不能有效抵抗脾切除后感染,组织学结构发生了质变.     

脾创伤保脾手术术式的选择（附158例分析体会）

目的　探讨脾创伤保脾术的术式选择。方法　对脾创伤采用术中保脾的 15 8例手术方式进行分析 ,其中氩气刀止血 5例 ,ZT生物胶止血 3例 ,单纯脾修补术 2 6例 ,脾部分切除术 77例 ,修补加脾部分切除术 12例 ,脾切除自体脾组织片网膜囊内移植术 35例。结果　全组病例治愈出院。脾切除自体脾组织片网膜囊内移植术组 35例中 ,2例出现粘连性肠梗阻 ,11例出现各类术后感染 ,而其它术中保脾组 12 3例中 ,10例出现术后感染 ,两组总感染数比较 ,P <0 .0 1。结论　脾创伤术中保脾术的术式选择 ,应根据病人个体情况及脾破裂的类型而定 ,必要时采用联合多种术式保脾。对伴有空腔脏器破裂者也可选择性保脾。但应慎重选择脾切除自体脾组织片网膜囊内移植术。     

自体脾移植在胃癌手术中的应用

目的 探讨对于胃癌患如何在行肿瘤根治术的同时，不失其脾脏的抗肿瘤免疫功能。方法 1991年1月至1994年7月间将39例Ⅲ期贲门胃体癌患随机分为3组：根治组(13例)行根治性全胃切除；扩大根治组(12例)行根治性全胃切除术加胰体尾和脾切除术；脾移植组(14例)行扩大根治术加自体脾移植。术后通过随访，进行前瞻性对比观察。结果 5年生存率：根治组为23．1％；扩大根治组为22．5％；脾移植组为50．0％。生存分析表明脾移植组5年生存率高于根治组(P=0．0463)和扩大根治组(P=0．0249)，后两组间差异无显性意义(P=0．6124)。术后两个月移植脾脏经^99mTc同位素扫描全部清晰显影即恢复功能。结论 自体脾组织移植能延长胃癌根治术后患的5年生存率。     

脾损伤自体脾组织移植的临床应用

目的：探讨自体脾组织移植在临床中的应用。方法：总结32例脾外伤行全脾切除自体脾组织移植手术，其中采用大网膜囊内移植18例，去粘膜游离空肠段内移植12例，腹直肌鞘内移植2例。结果：术后随访均显示脾功能满意，尤以去粘膜游离空肠段内移植效果最好。结论：自体脾组织移植可作为严重脾外伤、全脾切除术后保留脾功能的一个重要有效手段，移植脾的功能恢复与血供有密切的关系。     

自体移植脾巨噬细胞C_(3b)、Fc受体及蛋白质表达的实验研究

目的　探讨自体脾组织移植后的功能状况。方法　采用小鼠进行自体脾组织网膜内移植 ,术后 6个月切取移植脾组织 ,检测巨噬细胞的Fc、C3b受体及蛋白表达。结果　自体移植脾巨噬细胞Fc受体的含量与原位脾相近 ,C3b受体的功能正常 ;蛋白质的表达与原位脾相同。结论　大网膜内自体移植脾组织的功能在细胞水平是正常的 ,移植脾组织具有原位脾的功能。     

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.     

大网膜包裹人工神经移植体再血管化及神经再生的研究

目的 探讨大网膜包裹人工神经移植体的早期再血管化，以及增强移植体血供对神经再生的影响。方法 75只大耳白兔制作成左前肢正中神经2．0cm缺损，随机分成3组：A组，带蒂大网膜包裹人工神经移植体移植；B组，人工神经移植体桥接神经缺损；C组，自体神经桥接神经缺损，作为对照组。术后3、7和14d应用伊凡思蓝（evans blue bound to albumin，EBA）毛细血管造影，检测各组移植体的再血管化；术后12周通过电生理、光镜、透射电镜等检测评估神经再生的效果。结果 A、C组术后3d移植神经出现再血管化，7d和14d再血管化程度逐渐增强；B组7d出现再血管化，与A、C组比较，再血管化延迟。术后12周，A组和C组运动神经传导速度、有髓神经纤维密度、神经内纤维组织面积及面积比、髓鞘厚度、髓鞘直径等差异无统计学意义（P〉0．05），但均优于B组，差异有统计学意义（P〈0．05）。结论 大网膜包裹人工神经移植体可促进移植体早期再血管化，增强移植体血供，促进神经再生。     

Splenic tissue autotransplantation in rabbits: no restoration of host defense

BACKGROUND: The loss of spleen may increase the incidence of overwhelming sepsis. To prevent this, splenic autotransplantation has been performed in humans and experimental animals. However, there is still controversy about the effectiveness of regenerated splenic tissue in preventing infection. This study explored the effectiveness of splenic tissue autotransplantation in restoring host defense. MATERIALS AND METHODS: Rabbits were divided into three groups: splenic autotransplantation, sham operation, and total splenectomy. Histomorphology, T-lymphocyte count, serum lysozyme levels, hemolysin titers, and pneumococcal clearance were observed as read-out parameters over 24 weeks. RESULTS: Histological study showed that the white pulp was poorly developed and central arterioles were missing in the regenerated splenic tissue of the autotransplanted rabbits. The weight of regenerated spleens recovered 6 months later in the splenic autotransplantation group was 11% of that in the sham operation group and was significantly less than the weight at implantation. There was no significant difference in the number of T lymphocytes or level of serum lysozyme between the three groups. A poor antibody response by the rabbits in the splenic autotransplantation and total splenectomy groups was noted after the primary intravenous administration of sheep red blood cells compared to those of sham operation group. After the challenge with type 3 pneumococci intravenously, pneumococcal clearance from the bloodstream in the splenic autotransplantation group did not differ significantly from that in the total splenectomy group, but was markedly delayed compared with that in the sham operation group. CONCLUSIONS: The low quantity and poor quality of the regenerated splenic tissue contribute to the inferior immunoprotective ability of animals autotransplanted with one-third of the original spleen. This suggests that the regenerated spleen cannot compensate for the immunological function of the original one, especially host resistance to infection.     

急性出血坏死性胰腺炎大鼠胰腺组织结构改变与内毒素血症的关系及纳屈酮的治疗作用

为探讨大鼠急性出血坏死性胰腺炎（ＡＨＮＰ）胰腺组织结构改变与内毒素血症的关系及纳屈酮的治疗作用，用５％牛磺胆酸钠逆行胰胆管注射制成ＡＨＮＰ模型。取Ｗｉｓｔａｒ大鼠１１０只随机分为假手术组（ｎ＝２０）、ＡＨＮＰ组（ｎ＝４５）、纳屈酮治疗组（ｎ＝４５），分别于术后６、１２、２４小时称取胰腺重量，观察ＡＨＮＰ大鼠胰腺组织结构改变，同时测定其血浆淀粉酶和内毒素水平，并与假手术组比较。结果：与假手术组比较，ＡＨＮＰ组血浆淀粉酶、内毒素及胰腺系数升高，在光镜及电镜下可见胰腺损害随时间延长而加重。纳屈酮治疗组与ＡＨＮＰ组相比，其血浆淀粉酶、内毒素及胰腺系数下降，而且胰腺损害减轻。本实验结果提示：ＡＨＮＰ引发内毒素血症，纳屈酮可通过改善胰腺缺血性损害而降低血浆内毒素，延长大鼠生存时间及降低病死率     

Optimal site and amount of splenic tissue for autotransplantation.

Clinical and basic studies have documented a high susceptibility to pneumococcal infection in asplenic humans and animals. It has been suggested that autotransplantation of splenic tissue might be a method of providing host resistance when total splenectomy is necessary. However, the effect of splenic autograft has remained controversial. This study was performed to evaluate the most effective site and amount of splenic autograft using rats. Rats were divided into five groups for the purpose of determining the site of splenic autotransplantation: splenectomy, sham operation, implantation into the omental pouch, intraperitoneal implantation, and intramuscular implantation. For determining the amount for autotransplantation, the rats were divided into seven groups: splenectomy, sham operation, and implantations of 25, 50, 100, 200, or 300 mg of splenic tissue. All animals were challenged with Streptococcus pneumoniae type 6, 16 weeks after surgery. Howell-Jolly bodies appeared postsplenectomy, but disappeared in the implanted rats 16 weeks after the operation. Histologically, the implanted tissue was indistinguishable from that of a normal spleen. Pneumococcal clearance from the bloodstream and survival rate were significantly higher in rats implanted in the omental pouch as compared with splenectomized rats. Intraperitoneal and intramuscular implanted rats did not show a significant difference from the splenectomized rats. More than 50% of splenic tissue for autograft showed a significant increase in pneumococcal clearance and survival rate as compared with that of splenectomized rats. It was suggested that the most effective site of autotransplantation is the omental pouch and approximately 50% of the whole spleen would be necessary for prevention from sepsis.     

A Pilot Study: The Therapeutic Effect of Ultrasound following Partial Rupture of Achilles Tendons in Male Rats

The purpose of this pilot study was to determine whether ultrasound administered in clinical dosage has a therapeutic effect on the healing of partially ruptured Achilles tendons. The subjects were 11 young male Marland rats which were divided into two main groups. One group was treated for 2 weeks, the other for 3. Each group consisted of experimental and control animals, the control animals being mock-sonated only. A day after the final treatment the tensile strength of the Achilles tendon of all the rats was assessed. In addition, a light microscope and an electron microscope analysis of Achilles tendon tissue from rats on the 3 week treatment program was done. The statistical analysis of the data indicated a significant difference only between the experimental and control animals in the group sonated for 3 weeks. Electron microscope analysis showed ultrastructural changes in the tissue of the experimental animal consonant with those changes that would be found in a more advanced stage of the healing process.J Orthop Sports Phys Ther 1988;10(2):39-46.     

Electron microscopy of the exocrine pancreas in experimental acute hypercalcemia

Hypercalcemia has been associated with acute pancreatitis clinically and in the experimental animal. We studied the pancreatic ultrastructure in acute experimental hypercalcemia. Anesthetized cats (Pentobarbital, 0.55 mg/kg) received Ca++ (Calcium-Gluconate: 0.6 mmol/kgh; n = 4), K+ (KCl: 1.1 mmol/kgh; n = 4) or NaCl (0.9%; n = 4) locally through retrograde infusion into the splenic artery. Biopsies for electron microscopy (EM) were taken at three hours. Eight cats received intravenous Ca++ (0.6 mmol/kgh, 0.3 mmol/kgh after three hours) or NaCl (0.9%) for 12 hours. Biopsies were collected in two animals in three-hour intervals, and in all animals at twelve hours. After local calcium infusion necrotizing pancreatitis was seen macroscopically in the body of the pancreas. Biopsies for EM showed acinar cell necrosis, hydrops of nuclei and mitochondria and needle-like precipitates in the cytoplasma in the center of calcium perfusion. Biopsies taken from the peripheral region of the macroscopically altered tissue revealed desorganisation of the acinar polarisation and the endoplasmic reticulum, with zymogen granules appearing in the basolateral cell-portion. After intravenous calcium administration no macroscopical changes were seen. In EM acinar cells showed dilatation and proliferation of the golgi apparatus and increased number of condensing vacuoles indicating stimulation. Again, disorganisation of acinar cell polarisation was present. Control animals treated with K+ or NaCl showed normal pancreatic ultrastructure. The morphological changes after calcium infusion indicate direct damage to the acinar cell. Our results suggest that hypercalcemia induced pancreatitis could originate in the acinar cell.     

The effect of site and technique of splenic tissue reimplantation on pneumococcal clearance from the blood

The technique and site of reimplantation of splenic tissue influences survival of laboratory animals following intravenous injection of pneumococci. Splenic tissue was prepared by slicing, mincing, or grating the spleen. The tissue was placed subcutaneously, intraperitoneally, retroperitoneally, or in an omental pouch. This study was designed to determine the rate of pneumococcal clearance from the blood stream 16 weeks following splenic reimplantation by four different methods. All animals were challenged with an intravenous 1 mL bolus containing 10(7) bacteria. The New Zealand white rabbits were divided into six groups: intact spleen; splenectomized; spleen slices in an omental pouch; minced spleen in an omental pouch; splenic tissue implanted subcutaneously; and bits of spleen dropped into the peritoneal cavity. Animals with an intact spleen and those with spleen slices implanted into an omental pouch cleared bacteria during the first hour and all bacteria had disappeared at three hours. Bacteremia persisted longer than three hours in the other groups. Splenic tissue had regenerated in all animals with omental pouch implants, in four of six with minced spleen dropped into the peritoneal cavity but in only one with a subcutaneous implant. Reimplanted splenic tissue clears pneumococci from the blood stream best when thin slices of spleen are placed in an omental pouch. This technique also assures successful regeneration of splenic tissue.     

缺血后处理减轻犬肾缺血再灌注损伤的作用

目的 探讨缺血后处理减轻犬肾缺血再灌注损伤的作用及其相关机制.方法 随机将犬分为假手术组、缺血再灌注组和缺血后处理组,每组5只.假手术组:犬麻醉后,取其腹正中切口进入腹腔,游离双侧肾脏,切除右肾后,关腹.缺血再灌注组:手术操作与假手术组相同,仅在切除右肾和游离左肾之后,将左肾动、静脉夹闭60 min,然后开放血管.缺血后处理组:手术操作与缺血再灌注组相同,仅在肾动、静脉被夹闭60 min后,以再灌注(开放血管)30 s、夹闭血管30 s为1个循环,共进行6次循环,然后完全放开血管.分别于术后24、48及72 h采集犬静脉血2 ml,使用全自动生化分析仪测定各组犬血清肌酐(Cr)和尿素氮(BIJN)水平;术后第3天取犬肾组织,采用硫代巴比妥酸法测定丙二醛(MDA)含量,采用黄嘌呤氧化酶法测定超氧化物歧化酶(SOD)活性,采用化学比色法测定髓过氧化物酶(MPO)活性,并观察犬肾组织的病理改变和细胞凋亡情况.结果 术后各时间点,缺血再灌注组、缺血后处理组和假手术组犬的血清Cr和BUN水平均依次降低,3组间比较,差异均有统计学意义(P<0.05).术后第3天,缺血再灌注组、缺血后处理组和假手术组犬肾组织中SOD活性依次升高,而MDA含量和MPO活性均依次降低,3组间比较,差异均有统计学意义(P<0.05).假手术组肾小球和肾小管结构正常,未见明显病理改变;缺血再灌注组肾间质水肿,大量炎症细胞浸润,肾小管上皮细胞刷状缘消失,大量上皮细胞坏死、脱落,管腔扩张,其中可见大量管型;缺血后处理组可见肾间质轻度水肿,肾小管上皮细胞扁平,部分刷状缘消失、坏死,偶见管型,管周血管有少量淤血.假手术组、缺血再灌注组、缺血后处理组犬肾的细胞凋亡指数分别为2.7±1.3、28.4±6.2和15.4±4.1,3组间比较,差异均有统计学意义(P<0.05).结论 缺血后处理能减轻犬肾缺血再灌注损伤,其机制可能与缺血后处理减少氧自由基的产生、抑制细胞凋亡及减少炎症细胞浸润有关.     

Immunologic function against infection in splenic autotransplanted mice

The increasing recognition of the danger of overwhelming postsplenectomy infection (OPSI) has led surgeons to attempt to maintain splenic function after spleen injury. One technique they use when splenorrhapy or partial splenectomy are not feasible is the deliberate autotransplantation of splenic tissue. But the amount of splenic tissue necessary to prevent OPSI remains controversial, and opinions differ about the importance of the location and size of the splenic fragments implanted. The mice were divided into five groups, I. splenectomy, II. splenectomy +30% of the spleen implanted intraperitoneal site, III. splenectomy +50% implanted intraperitoneally, IV. splenectomy +50% implanted subcutaneously and V. Sham operation. This study assessed the blood flow of the splenic tissue, increasing weight of splenic mass, histology, the serum level of the immunoglobulins (IgG, IgA, and IgM), pneumococcal antibody titers after vaccination, and survival after intravenous pneumococcal challenge. This study demonstrated that intraperitoneal transplantation showed better regeneration and afforded better protection from OPSI than subcutaneous transplantation. And 30 to 50 percent of the whole splenic tissue mass protected against experimental pneumococcal sepsis. The splenic autotransplants developed in volume and blood supply after 8 weeks, and immunologic function against infection recovered at the same time.     

预防创伤后膝关节粘连的实验研究

目的　探讨多聚合高分子纤维素 (POLYCMC)预防外伤后关节粘连的作用。方法　实验用兔 4 8只 ,动物随机分成三组 ,多聚合高分子纤维素组 (A组 ) ,地塞米松组 (B组 ) ,对照组 (C组 )。术后下肢管型石膏固定 ,各组分别于术后 3周 ,6周处死 ,进行生物力学测定 ,肉眼观察 ,光镜及透射电镜观察 ,并对有关数据进行相应的统计学分析。结果　在膝关节活动度方面 ,A组与B组和C组比较均有高度显著的统计学差异 (P <0 0 1 ) ,即在膝关节活动度方面 ,A组优于B组及C组。在大体标本粘连等级方面 ,A组与B组和C组比较均有高度显著的统计学差异 (P <0 0 1 ) ,即A组粘连程度轻于B组及C组。光镜、透射电镜的组织学观察显示A组滑膜上皮细胞增生不明显 ,纤维结缔组织少 ,细胞形态接近正常 ,基质胶原原纤维少。结论　多聚合高分子纤维素可以有效地预防外伤后膝关节粘连