兔肝细胞的脾脏移植和定位初步研究

【目的】探讨肝细胞体内移植的途径和定位分析。【方法】从新西兰白兔的肝脏中分离获得原代肝细胞，经纯化培养后经脾动脉插管或注射移植到脾脏，在不同时间注射肝细胞特异性增强剂（扎贝葡胺），观察移植后脾脏核磁信号的变化和移植肝细胞的存活，并分析细胞移植前后肝功能变化。【结果】肝细胞移植入脾脏一段时间后受损肝功能明显改善。脾脏内可见肝细胞信号，d，信号最强，以后逐渐衰退并持续14d以上。【结论】肝细胞脾内移植是可行的；应用肝细胞增强剂进行MRl分析可成为肝细胞移植无创伤检测手段。

移植脾组织血管神经生成规律的动态观察

目的:观察自体脾组织大网膜内移植后,移植脾组织血管神经再生过程中血管内皮细胞生长因子、血管内皮细胞生长因子受体及神经肽Y表达的动态变化,分析自体移植脾组织血管神经生成规律。方法:实验于2004-09/2006-10在解放军第三军医大学基础部外科应用解剖与手术学教研室实验室完成。①选用Wistar大鼠70只,按随机数字表法分为7个时相组,每组10只。②行脾切除自体脾组织大网膜内移植,分别于术后7,14,30,60,90,120,180d,采用免疫组化血管内皮细胞生长因子、血管内皮细胞生长因子受体、神经肽Y阳性神经纤维抗体染色方法,应用光镜、电镜、图像分析观测自体移植脾组织。结果:70只大鼠全部进入结果分析,无脱失。①自体脾组织移植术后7d即有血管长入移植脾组织,180d再生血管接近正常;术后30d神经开始再生,180d趋于正常。②术后7,14d血管内皮细胞生长因子、血管内皮细胞生长因子受体阳性染色细胞密度迅速升高,60d达高峰,随后逐渐降低;术后30d出现神经肽Y阳性神经纤维。术后180d血管内皮细胞生长因子、血管内皮细胞生长因子受体阳性染色细胞密度趋于正常;术后120,180d神经肽Y阳性神经纤维广泛分布。结论:自体移植脾组织再生过程中移植脾组织再生血管神经来源于大网膜内的神经。自体移植再生脾组织内血管内皮细胞生长因子、血管内皮细胞生长因子受体表达量早期开始升高,移植脾组织先出现血管再生,后出现神经再生。

自体脾移植治疗小儿外伤性脾破裂23例报道

目的探讨自体脾移植在小儿外伤性脾破裂全脾切除术中的疗效。方法对本院近10年来收治的23例小儿外伤性脾破裂患儿行全脾切除+自体脾组织大网膜内移植术,评价其疗效。结果本组均治愈。随访20例,随访期患儿生长、发育正常,IgM、IgG在正常范围,无脾切除后暴发性感染(OPSI)发生。结论全脾切除+自体脾组织移植治疗小儿外伤性Ⅲ级、Ⅳ级脾破裂能降低死亡率,保留脾脏的大部分功能。

自体脾移植对免疫功能的影响

为探讨外伤性脾破裂行脾切除术 (SE )与脾切除术加自体脾组织移植 (AS)对免疫功能的影响。笔者观察了 64例SE和 42例AS的术后感染性疾病发生率与血清免疫物质水平的关系。结果显示 :SE的感染率为 78.1 3 % ,AS为 61 .90 % ,差别无显著性意义 (P >0 .0 5)。SE的IgG和IgA高于AS(P <0 .0 5) ,AS高于对照组 (P <0 .0 5) ,SE的IgM低于AS(P >0 .0 5) ,显著低于对照组 (P <0 .0 5)。C3的水平三者差别均无显著意义 (P >0 .0 5)。提示 :自体脾片移植能保留脾切除术后的部分免疫功能。

自体脾组织移植在外伤性脾破裂手术中的应用

【目的】探讨自体脾组织移植在外伤性脾破裂治疗中的应用,评价其疗效。【方法】回顾性分析本院60例外伤性脾破裂患者的临床资料,其中A组26例行脾大部分或全脾切除术,另B组34例行脾大部分或全脾切除术加自体脾组织移植术;比较两组患者术后住院时间、术后感染、术后腹腔引流量、血浆免疫球蛋白和Tuftsin蛋白浓度的差异。【结果】两组患者术后住院时间、血浆免疫球蛋白浓度无明显差异(P>0.05);而术后感染、腹腔引流量与血浆TuFtsin蛋白浓度均有明显差异,A组明显优于B组(P<0.05)。【结论】脾切除加自体脾组织移植术能明显减少术后感染,是治疗严重外伤性脾破裂较为有效的手术方法。

非酒精性脂肪性肝病临床分析及海狗油疗效观察

目的对照性分析非酒精性脂肪性肝病(NAFLD)临床特点,探讨海狗油ω-3多不饱和脂肪酸胶丸(海狗油)对NAFLD的治疗效果。方法①收集NAFLD患者和正常健康人资料各76例,以病例对照研究方法分析NAFLD的临床生化指标。②选取18例NAFLD患者,予以海狗油连续治疗12周,动态观察海狗油对NAFLD患者临床生化和影像学检查结果的影响。结果①NAFLD组体重指数(BMI)、谷丙转氨酶(ALT)、甘油三酯(TG)、极低密度脂蛋白胆固醇(VLDL-C)、空腹血糖(FPG)显著高于对照组(P<0.01),两组间AST和TCH差异无显著性。②NAFLD组合并肥胖、高血压、血脂紊乱、高血糖中两项或两项以上者和代谢综合征者分别为72.37%(55/76)和46.05%(35/76)。③与治疗前比较,海狗油治疗4周末和8周末时,NAFLD患者的血清LP-a显著降低(P<0.01);治疗12周末时,血清Apo-A1明显降低(P<0.05),肝/脾CT值比值的差异无显著性(P>0.05)。结论NAFLD与肥胖、血脂紊乱和高血糖密切相关,海狗油对NAFLD患者的血脂有一定调节作用。

肝卵圆细胞经脾脏移植治疗肝纤维化的实验研究

目的研究肝卵圆细胞在治疗肝纤维化的有效性。方法用携带GFP转基因C57BL／6小鼠，喂0．1％的3，5-二乙酯基-1，4二氢三甲基吡啶（DDC），刺激卵圆细胞增生，再两步酶灌注法和Sea-1抗体标记的免疫磁珠分选法加以分离出携带有GFP的SCa-1＋细胞。利用皮下注射CCI。的方法，建立小鼠肝纤维模型。根据不同的组别经脾分别输入卵圆细胞或生理盐水。4周后观察肝功能各项指标及肝脏的病理改变，比较各组间的差别。结果经脾脏移植后卵圆细胞入肝后主要定植与汇管区。移植后肝功能指标均有不同程度改善，纤维化程度有所减轻，尤以停止注射四氯化碳组明显。结论携带GFP的基因卵圆细胞经脾注射至肝纤维化小鼠体内后，可定植于受者肝脏内，并可缓解肝硬化的进展，促进肝纤维的逆转。

肝卵圆细胞与骨髓间质干细胞经脾移植治疗实验性小鼠肝纤维化的疗效比较

目的比较肝卵圆细胞（HOC）与骨髓间充质干细胞（MSC）治疗小鼠肝纤维化的疗效。方法以含浓度为0．1％的3，5-二乙酯基．1，4二氢三甲基吡啶（DDC）喂养C57BL／6小鼠，建立小鼠肝卵圆细胞增殖模型，而后利用胶原酶原位灌注及percoll密度梯度离心法分离肝脏非实质细胞，再用免疫磁珠分离纯化出Sca-1阳性的肝卵圆细胞。利用四氯化碳（CCl4）皮下注射诱导小鼠肝纤维化模型。最后经脾分别输入卵圆细胞、骨髓间充质干细胞或生理盐水（NS）。4周后观察肝功能各项指标、羟脯氨酸及肝脏的病理改变，比较各组间的差别。结果在继续注射CCl4的情况下，HOC移植与MSC移植治疗均能改善肝纤维化小鼠的肝功能，降低肝脏羟脯氨酸含量，缓解肝纤维化，但HOC移植组更加明显（P〈0．05）；在停用CCl4后，各组肝纤维化均有不同程度缓解，差异不明显（P〉0．05）。结论肝卵圆细胞经脾注射移植后，能改善肝硬化小鼠的肝功能，减轻其肝纤维化的程度。且效果比骨髓间充质干细胞更明显。

Diminution of toxic copper accumulation in toxic milk mice modeling Wilson disease by embryonic hepatocyte intrasplenic transplantation

AIM: To observe the therapeutic effect of intrasplenic transplantation with embryonic hepatocytes on amelioration of hereditary copper accumulation in toxic milk (TX) mouse modeling Wilson disease. METHODS: Donor hepatocytes were harvested from 14-d fetal liver of a pregnant homogeneous DL mouse. These cells were successively cultured, labeled with fluorescein dye Hoechst 33342 for 24 h, and sequentially infused into the spleen parenchyma of the recipient TX mice. No host immunosuppression measures were taken. Two and four weeks after transplantation, the recipients were killed for routine histologic investigation and immunohistochemistry study up to 4 wk after transplantation. The serum copper and ceruloplasmin concentrations of the recipient mice were determined by graphite furnace atomic absorption spectroscopy.RESULTS: In the following 2nd and 4th wk after transplantation, the donor hepatocytes could be visualized in the livers of 47.3% recipients. The serum ceruloplasmin and copper concentrations increased by 1.6-fold after 2 wk and 2.0-fold times after 4 wk respectively, which ultimately rose from about 30% of the normal level to nearly 60%(P＜0.01). The hepatic copper concentration decreased 7.2%, 4 wk after transplantation. Pathologic examination showed that there were many actively proliferative hepatocyte precursor cells with specific embryonic hepatocyte marker AFP migrated into hepatic sinusoidsof the recipients. A large number of cells carrying hepatocytes marker and albumin were observed in the recipient spleen tissues.CONCLUSION: Embryonic hepatocytes are capable of differentiating into mature hepatocytes in vivo. After transplantation, the hereditary abnormalities of copper metabolism in TX mice could be corrected partially by intrasplenic transplantation of homogeneous embryonic hepatocytes.

Embryonic stem cells develop into hepatocytes after intrasplenic transplantation in CCl_4-treated mice

AIM： To transplant undifferentiated embryonic stem （ES） cells into the spleens of carbon tetrachloride （CCl4）-treated mice to determine their ability to differentiate into hepatocytes in the liver. METHODS： CCh, 0.5 mL/kg body weight, was injected into the peritoneum of C57BL/6 mice twice a week for 5 wk. In group 1 （n = 12）, 1 × 10^5 undifferentiated ES cells （0.1 mL of 1 × 10^6/mL solution）, genetically labeled with GFP, were transplanted into the spleens 1 d after the second injection. Group 2 mice （n = 12） were injected with 0.2 mL of saline twice a week, instead of CCh, and the same amount of ES cells was transplanted into the spleens. Group 3 mice （n = 6） were treated with CCh and injected with 0.1 mL of saline into the spleen, instead of ES cells. Histochemical analyses of the livers were performed on post-transplantation d （PD） 10, 20, and 30. RESULTS： Considerable numbers of GFP-immunopositive cells were found in the periportal regions in group 1 mice （CCh-treated） on PD 10, however, not in those untreated with CCh （group 2）. The GFP-positive cells were also immunopositive for albumin （ALB）, alpha-1 antitrypsin, cytokeratin 18, and hepatocyte nuclear factor 4 alpha on PD 20. Interestingly, most of the GFP-positive cells were immunopositive for DLK, a hepatoblast marker, on PD 10. Although very few ES-derived cells were demonstrated immunohistologically in the livers of group 1 mice on PD 30, improvements in liver fibrosis were observed. Unexpectedly, liver tumor formation was not observed in any of the mice that received ES cell transplantation during the experimental period CONCLUSION： Undifferentiated ES cells developed into hepatocyte-like cells with appropriate integration into tissue, without uncontrolled cell growth.

Effects of NGF and TrkA on GAP-43^+ nerve regeneration in rat autotransplanted splenic tissue

Objective:To study the time-course of the regeneration of GAP-43+ nerve, and the effects of NGF and TrkA on this process. Methods: Adult Wistar rats underwent splenectomy and splenic autotransplantation, or sham-operation. On day 7, 14, 30, 60, 90, 120, and 180 after surgery, the density of GAP-43+ nerve fibers in spleen tissues were measured with the immunohistochemistry followed by computer image analysis. The expressions of GAP-43, NGF and TrkA were determined with in situ hybrdization, and their mRNA levels were detected with RT-PCR and image analysis qualification. Results: (1) The GAP-43+ nerve fibers began their regeneration on 30 d after operation and extended from greater omentum into splenic autotransplants. Density of the nerve fibers gradually became greater and almost normal 180 d after operation. (2) In splenic autografts, the mRNA expression of GAP-43, NGF and TrkA appeared on day 30 after the operation, gradually reached the peak on day 90. Conclusion: The renascent GAP-43+ nerve fibers may come from the greater omentum packaging the splenic autografts and NGF and TrkA can promote the nerval regeneration in the autotransplant spleen tissues.

Formation of microchimerism in rat small bowel transplantation by spienocyte infusion

AIM： To investigate the effect of donor splenocyte infusion combined with cyclosporine A （CsA） on rejection of rat small bowel transplantation （SBT）. METHODS： Male Sprague-Dawley （SD） rats and female Wistar rats weighing 230-270 g were used as donors and recipients respectively in the study. Heterotopic small bowel transplantation was performed. The rats were divided into three groups： group one receiving allotransplantation （SD→Wistar）, group two receiving allotransplantation （SD→Wistar） ＋ donor splenocyte infusion, group three receiving allotransplantation （SD →Wistar） ＋ donor splenocyte infusion ＋ CsA followed by CsA 10 mg/kg per day after transplantation, in which recipient Wistar rats were injected with 2 ×10^8 SD splenocytes 28 d before transplantation, and treated with CsA after transplantation. Finally, the specific DNA fragment of donor Y chromosome was detected in recipient peripheral blood and skin by PCR. The survival time after small bowel transplantation was observed. Gross and histopathological examinations were performed. RESULTS： The survival time after small bowel transplantation was 7.1 ± 1.2 d in group 1, 18.4 ± 3.6 d in group 2 and 31.5± 3.1 d in group 3. The survival time was significant longer （P 〈 0.01） in group 3 than in groups 1 and 2. The gross and histopathological examination showed that the rejection degree in group 3 was lower than that in groups 1 and 2.CONCLUSION： Donor splenocyte infusion combined with CsA decreases remarkably the rejection and prolongs the survival time after rat small bowel transplantation.

Treatment modalities for hypersplenism in liver transplant recipients with recurrent hepatitis C

Hepatitis C is the most common indication for orthotopic liver transplantation in the United States. Unfortunately, hepatitis C recurs universally in the transplanted liver and is the major cause of decreased graft and patient survival. The combination therapy of interferon and ribavirin has been shown to be the most effective therapy for recurrent hepatitis C. However, pre-and post-transplant hypersplenism often precludes patients from receiving the antiviral therapy. Splenectomy and partial splenic embolization are the two invasive modalities that can correct the cytopenia associated with hypersplenism. In this report we review the two treatment options, their associated outcomes and complications.

Management of carbon tetrachloride-induced acute liver injury in rats by syngeneic hepatocyte transplantation in spleen and peritoneal cavity

AIM:Acute hepatitis may seldom have a fulminant course. In the treatment of this medical emergency,potential liver support measure must provide immediate and sufficient assistance to the hepatic function.The goal of our study was to study the adequacy of hepatocyte transplantation (HCTx)in two different anatomical sites,splenic parenchyma and peritoneal cavity,in a rat model of reversible acute hepatitis induced by carbon tetrachloride(CCl_4). METHODS:After CCl_4 intoxication,84 male Wistar rats used as recipients were divided in to four experimental groups accordingly to their treatment:Group A(n=-24):intrasplenic transplantation of 10×10~6 isolated hepatocytes,Group B(n=24): intraperitoneal transplantation of 20×10~6 isolated hepatocytes attached on plastic microcarriers,Group C(n=-18):intrasplenic injection of 1 mL normal saline(sham-operated controls), Group D(n=-18):intraperitoneal injection of 2.5 mL normal saline(sham-operated controls).Survival,liver function tests (LFT)and histology were studied in all four groups,on d 2, 5 and 10 post-HCTx. RESULTS:The ten-day survival(and mean survival)in the 4 groups was 72.2%(8.1±3.1),33.3%(5.4±3.4),0% (3.1±1.3)and 33.3%(5.4±3.6)in groups A,B,C,D, respectively(P_(AB0<0.05,P_(AC)<0.05,P_(BD)=NS).In the final survivors,LFT(except alkaline phosphatase)and hepatic histology returned to normal,independently of their previous therapy.Viable hepatocytes were identified within splenic parenchyma(in group A on d 2)and both in the native liver and the fatty tissue of abdominal wall(in group B on d 5). CONCLUSION:A significantly better survival of the intrasplenically transplanted animals has been demonstrated. Intraperitoneal hepatocytes failed to promptly engraft.A different timing between liver injury and intraperitoneal HCTx may give better results and merits further investigation.

Successful laparoscopic splenectomy after living-donor liver transplantation for thrombocytopenia caused by antiviral therapy

Although interferon (IFN) based therapy for recurrent hepatitis C virus (HCV) infection after liver transplantation has been widely accepted, it induces various adverse effects such as thrombocytopenia, resulting in its interruption. Recently, concomitant splenectomy at the time of living donor liver transplantation (LDLT) has been tried to overcome this problem, but this procedure leads to several complications such as excessive intraoperative bleeding and serious infection. A 60-year-old female received LDLT using a left lobe graft from her second son for liver failure caused by hepatitis C-related cirrhosis. Six months after LDLT, she was diagnosed as recurrent HCV infection by liver biopsy. IFN monotherapy was started from 7 mo after LDLT and her platelet count decreased to less than 50 000/μL, which thus made it necessary to discontinue the treatment. We decided to attempt laparoscopic splenectomy (LS) under general anesthesia. Since intra-abdominal findings did not show any adhesion formations around the spleen, LS could be successfully performed. After LS, since her platelet count immediately increased to 225 000/μL 14 d after operation, IFN therapy was restarted and we could convert the combination therapy of IFN and ribavirin, resulting in no detectable viral marker. Inconclusion, LS can be performed safely even after LDLT, and LS after LDLT is a feasible and less invasive modality for thrombocytopenia caused by antiviral therapy.

Severe thrombocytopenia before liver transplantation is associated with delayed recovery of thrombocytopenia regardless of donor type

AIM： To compare the recovery of thrombocytopenia and splenomegaly during long-term follow-up after liver transplantation in patients receiving a living donor transplant or a cadaveric donor transplant. METHODS： This was a retrospective cohort study of 216 consecutive liver transplant patients who survived for 〉 6 mo after transplantation; 169 received a liver transplant from a living donor and 47 from a cadaveric donor. The platelet counts or spleen volumes were examined before transplant, i, 6, and 12 mo after transplant, and then annually until 5 years after transplant. RESULTS： The mean follow-up period was 49 mo （range, 21-66）. Platelet counts increased continuously for 5 years after orthotopic liver transplant. The restoration of platelet counts after transplant was significantly slower in patients with severe pretransplant thrombocytopenia （〈 50000/μL） until 4 years after transplant （P = 0.005）. Donor type did not significantlyaffect the recovery of platelet count and spleen volume in either patient group. In multivariate analysis, pretransplant severe thrombocytopenia （〈 50000/μL） was an independent factor associated with sustained thrombocytopenia （P 〈 0.001, odds ratio 6.314; confidence interval, 2.828-14.095）. Thrombocytopenia reappeared after transplant in seven patients with portal flow disturbance near the anastomosis site. CONCLUSION： Our study suggests that severe thrombocytopenia before transplant is closely associated with delayed recovery of platelet count after transplant and donor type did not affect the recovery of thrombocytopenia. The reappearance of thrombocytopenia after transplant should be considered a possible indicator of flow disturbance in the portal vein.

EXPERIMENTAL STUDY ON THE SEGMENTAL PERFUSION AND PRESERVATION OF SPLEEN

Objective. Spleen transplanlation has developed to be an effective strategy for hemophilia A. But it has not been reported up to date that which kind of established solutions is most suitable for perfusion and preservation of spleen. This study aimed to establish some experiences with the comparison among Hartmann’s solution, Collins’ solution and WMO I solution, in order to instruct the clinical spleen transplantation. Methods. After the splenic artery and vein were dissociated clearly, three kinds of perfusion solutions began to perfuse the corresponding segments of spleen with a randomized sequence. When the efferent fluids from the splenic vein became clear, the perfused spleen segments were preserved for different durations with the same perfusing solution to calculate the survival rate of splencocyte(SRS) and were examined with light and electron microscopy. Results. Among the three solutions, SRS with WMO I solution was significantly higher than those of the other two(P< 0.001). The perfused spleen with WMO I solution showed the slightest morphological changes and a significant longer preservation duration than those with the other two(P<0.05 or P< 0 01). Conclusion. Among the three solutions, WMO I solution was most suitable for perfusion and preservation of spleen.

Changes of duplex parameters and splenic size in liver transplant recipients during a long period of observation

AIM: To assess the changes of portal and arterial velocities, resistance index, spleen and liver size during a long observation period (13.7 years) after orthotopic liver transplantation (OLT).METHODS: Two hundred and sixty patients were recruited retrospectively for this study and divided into groups with defined time intervals after OLT. The cross-sectional changes of portal and arterial velocities,resistance index, spleen and liver size between the defined time intervals were studied. The complications detected by ultrasound were compared to gold standard methods.RESULTS: The mean values for liver size were all within the normal range. The splenic size decreased between the time intervals 100 and 1 000 d after OLT (t;P＜0.01).While portal and arterial flow velocities decreased up to 5.5 years (t; portal velocity P＜0.01, maximal systolic velocity P=0.05, maximal end diastolic velocity P＜0.01),RI increased during this interval (t:P＜0.01). Higher RIvalues were found in older patients (r = 0.24, P＜0.001).CONCLUSION: The arterial and portal velocities show adaptation processes continuing over the course of many years after OLT and are reported for the first time. The vascular complications detected by ultrasound occur mostly up to 100 d after OLT.

Effect of operation-synchronizing transfusion of apoptotic spleen cells from donor rats on acute rejection of recipient rats after liver transplantation

AIM: To study effect of operation-synchronizing transfusion of apoptotic spleen cells from donor rats on acute rejection of recipient rats after liver transplantation. METHODS: Two of Wistar rats were chosen randomly for normal liver pathology control and ten of SD rats chosen randomly for liver function control as blank group (no operation). The rest of Wistar and SD rats were divided into four groups: control group (only liver transplantation), Dex group (donors receiving intraperitoneal injection of dexamethasone), SpC group (recipients receiving infusion of spleen cells of donors), Dex-SpC group (recipients receiving infusion of apoptotic spleen cells of donors), with each group except blank group, containing 10 SD rats and 10 Wistar rats, respectively. Wistar rats received liver transplantation from SD rats, in the meantime they received infusion of spleen cells of donors, which were induced by an intraperitoneal injection of dexamethasone (3 mg/(d.kg)·b.w) for three days before liver transplantation. The serum alanine transaminase (ALT), total bilirubin (T bili), liver pathological changes and survival time were analysed. Statistical analysis was carried out using SPSS 10.0 for Windows. Differences of the parametric data of ALT in means were examined by one-way ANOVA. Differences of ALT between two groups were examined by LSD. Differences of the nonparametric data of T bili in means and scores of pathology classification for acute rejection were examined by Kruskal-Willis H test. The correlations between ALT and T bili were analysed by Bivariate. Kaplan-Meier curves were used to demonstrate survival distribution. The log-rank test was used to compare the survival data. RESULTS: There were significant differences in ALT of the five groups (F= 23.164 P= 0.000), and ALT in Dex-SpC group was significantly higher than that in blank control, control, Dex, and SpC groups (P = 0.000), and ALT in SpC group was significantly higher than that in blank control (P= 0.000), control (P= 0.004), and Dex groups (P= 0.02). Results of nonparametric analysis of T bill showed that there were differences in T bill of the five groups (X2= 33.265 P= 0.000). T bili in Dex-SpC group was significantly higher than that in blank control, control, Dex, and SpC groups. T bili in SpC group was higher than that in blank control, control, and Dex groups. There were significant differences in scores of pathology classification for acute rejection in each of the groups (X2= 25.933, P= 0.000). The pathologically more serious acute rejection was found in Dex-SPC group than in other groups. No sign of acute rejection was observed in the blank control group. Slight acute rejection was observed in the control group. Slight-moderate acute rejection was observed in the Dex group. Moderate-acute rejection was observed in the SpC group. Severe-acute rejection was observed in the Dex-SpC group. The survival time in Dex-SpC group was shorter than in other groups (statistic = 11.13, P= 0.011). ALT and T bili were positively correlated (r= 0.747, P= 0.000, two-tailed). CONCLUSION: In order to reduce quantity of blood loss from rats after liver transplantation, only one of ALT or T bili is needed for liver function measurement of rats. Simultaneous injection of apoptotic spleen cells from donors induced by dexamethasone to liver transplantation rats aggravates acute rejection. One important mechanism of aggravation of acute rejection may be that apoptotic cells are not removed in time and that dead cells including apoptotic cells release inflammatory factors.

Determination of phagocytic functions of macrophages, vascular density and NPY-nerve within autotransplanted splenic tissue in adult rats

Objective: To estimate directly phagocytic functions of the macrophages because of the importance in innate immunity, determine blood vessel density and re-innervation density which are basis of function. Methods: Eighty adult Wistar rats were randonjy divided into experimental and control groups.The formers underwent splenotomy and a half splenie slice was transplanted into greater omentum. The latter only moved. After 6 months, examination was made as follows: ① After injection of 0.4% carbon particles by vein, spleme tissues were taken out at different times for estimating phagocytosis by light microscope. ② When splenic tissues had been intubated into left ventricle under total anesthesia, animals were perfused by formalin and India ink mixture suspension. Splenic tissues were taken out for making sections for measurement of area density of blood vessels.③ Immunohistochemical procedure was used for detecting neuropeptide Y(NPY). Results: Phagocytie functions had no difference between two groups, hut the area density of blood vessels and NPY-positive fibers re-dued (P<0.01) in experimental group. Conclusion:Autotransplanted splenic tissues show good innate immunity though regeneration of blood vessels and nerves do not reach normal level.