Liver transplantation in hamster-to-rat is a good model for the research in xenograft, but it is difficult to be performed. In order to simplify its procedures, 38 cases of liver transplantation in hamster-to-rat were performed with several technique improvments on the basis of orthotopic liver transplantation in rats. In the light of hamster’s anatomy, three cuffs anastomosis method was used. Because of its high stability and high survival rate, the model can be used widely as the research for liver xenograft.
摘要:目的：探讨成都地区体检人群中丙氨酸氨基转移酶（ALT）升高率与其升高的相关因素,为正确分析引起ALT升高的原因提供相关依据。方法:以参与体检的8734名体检人群为研究对象,收集身高、体重、血压、丙氨酸氨基转移酶、空腹血糖、高密度脂蛋白、低密度脂蛋白、总胆固醇、甘油三酯、血清HBsAg、脂肪肝及胆石症等相关资料进行分析。结果:在全部体检人群中,ALT升高率为1011%,男性ALT升高率为13.70%,女性ALT升高率为6.30%,男性明显高于女性（Plt;0001）；ALT升高组的年龄均数小于ALT正常组（Plt;0001）；在ALT升高的受检者中,脂肪肝、高脂血症、肥胖、糖尿病、胆囊结石、饮酒及乙肝等患病率均高于ALT正常组受检者（Plt;005）。结论:脂肪肝、糖脂代谢紊乱及乙肝是体检人员ALT升高的主要原因；男性和低龄也是体检者ALT升高的危险因素。Abstract: Objective: To investigate the prevalence and relative factors of elevated serum alanine aminotransferase(ALT) levels and providescientific bases for its causes analysis in physical examination people in Chengdu. Methods: Subjects who received medical examination in physical examination center of west China hospital were screened in this study. The information of height, body weight, blood pressure, serum ALT, fasting plasma glucose, highdensity lipoprotein cholesterol, lowdensity lipoprotein cholesterol, total cholesterol, triglyceride, hepatitis B surface antigen (HBsAg) statue, fatty liver and cholelithiasis were collected and analyzed. 〖WT5”HZ〗Results:〖WT5”BZ〗 A total of 8734 cases were included in this study. The total prevalence of elevated ALT was observed in 1011%, including 137% in man and 63% in woman, and this difference between man and woman was statistic significant (P＜0001). The mean age of ALT elevated group was obvious lower than that of normal ALT group (P＜0001). Interesting, the occurrence rates of fatty liver, hyperlipidemia, obesity, diabetes,gallstones, drinking and positive hepatitis B surface antigen in ALT elevated group were all significant higher than that in normal ALT group (P＜005). Conclusion: Fatty liver, glyeolipid metabolism disorder, and hepatitis B were main reasons of elevated ALT. Male and young cases were both high risk of elevated ALT in this study.
ObjectiveTo study the epidemiologic characteristics of primary liver cancer (PLC). MethodsThe literatures about regional distribution and etiologic epidemiology of PLC were reviewed. Results PLC was mainly distributed on caostland in the south-east of China. The main cause of PLC was hepatitis B virus, aflatoxin and contamination of drinking water. Otherwise, PLS was also related with lack of some trace element, sex horemones, genealogy cause and so on.Conclusion The genesis of PLC was by multiple factors.
ObjectiveTo analyze the death causes of postoperative early-stage after orthotopic liver transplantation (OLT) in rats, and to provide appropriate treatment strategies. MethodsThree hundreds of rat OLT models were performed by modified Kamada two-cuff technique. Operation time of each stage during OLT and postoperative survival time of rats were recorded and analyzed. According to survival time, the rats were divide into 4 groups:intraoperative death group (rats died during operation), < 6 hours group, 6-24 hours group, and > 24 hours group. Then comparison of operation time of each stage during OLT in rats of 4 groups was performed, and reasons of death during 24 hours after OLT were analyzed. ResultsOf the 300 OLT models, 37 rats died during operation (12.33%), 51 rats died within 6 hours after operation (17.00%), 76 rats died during 6-24 hours after operation (25.33%), and 136 rats survived longer than 24 hours (45.34%). The most common death causes of < 6 hours group were as follows:lose too much blood during the operation (27.45%, 14/51), postoperative bleeding (27.45%, 14/51), and vascular embolization (15.69%, 8/51). However, the most common death causes of 6-24 hours group were as follows:angiostenosis (27.63%, 21/76), postoperative bleeding (21.05%, 16/76), and pulmonary edema (19.74%, 15/76). There were significant differences in the cold ischemia time and anhepatic phase among the 4 groups (P < 0.05). The cold ischemia time and anhepatic phase of intraoperative death group were longer than those of other 3 groups (P < 0.05), in addition, the cold ischemia time of > 24 hours group was shorter than those of other 3 groups (P < 0.05). ConclusionsThere are many reasons leading to the early death after OLT. The long time of anhepatic phase and the cold ischemia time, intraoperative and postoperative bleeding, thrombosis, angiostenosis, and pulmonary edema are key factors for the improvement of prognosis in rats after OLT operation. Improvements of the reasons above are helpful to improve the successful rate of modeling and quality of OLT rats.
The effects on rat’s liver preservation using HX solation with high potassium and low sodium or HXm solution with high sodium and low potassium were studied with isolated perfusion of rat livers (IPRL). Sixty inbred Wistar rats were randomly divided into group HX (preserved with HX solution, n=30) and group HXm (preserved with HXm solution,n=30). The preservation effects of the storage solutions were assessed by measuring the sinunoidal lining cell mortality (SLCM), the Krebs-hense-leit perfusate ketone bidy ratio (PKBR), the hepatic sugar release (SL), and the hepatic tissue water content (HTWC). The results showed that there no significant differences between the two storage solutions after 6 hours preservation. If the preserved time was prolonged to 12 hours or more, the effect of rat’s liver preservation using HX solution were much superior to those using HXmsolutin.
Objective To study the culture and purification of the fetal mouse liver mesenchymal stem cells(MSCs) in vitro and to investigate their differentiation potential and the composite ability with true bone ceramic(TBC). Methods The single cell suspension of MSCs was primarily cultured and passaged, which was prepared from the fetal mouse liver; the flow cytometry was applied to detectCD29, CD34, CD44 and CD45. The osteogenic differentiation was induced in chemical inducing system; the osteogenic induction potency was tested. The purified fetal mouse liver MSCs were compounded with TBC covered with collagen type Ⅰ in vitro and the cell attachment and proliferation to the TBC were observed. Results The primary MSCs of fetal mouse liver were easy to culture in vitro. They proliferated well and were easy to subcultured. The proliferation ability of primary and passaged MSCs was similar. Flow cytometric analysis showed the positive results for CD29, CD44 and the negative results for CD34, CD45. After 7 days of induction, the MSCs expressed collagen type I and alkaline phosphatase(ALP) highly. After 14 days of induction, the fixed quantity of ALP increased significantly. After 28 days of induction, calcium accumulation was observed by Von Kossa’s staining. Many liver MSCs attached to the surface of TBC. Conclusion The MSCs of the fetalmouse liver can be obtained, subcultured and purified easily. After culturing in chemical inducing system, the MSCs of fetal mouse liver can be successfully induced to osteoblast-like cells, attach to the surface of TBC and proliferate well.
Objective To investigate the significance of three dimensional (3D) visualization and virtual surgery system in liver transplantation surgery. Methods Two patients suffered from cholangiolithiasis were scanned by 64 slice helical-CT on livers and the data were collected. Man-made segmentation and true up on the image from the data were carried out. 3D moulds of the liver and the intrahepatic vessels were reconstructed by VTK software respectively. And then, the moulds were imported to the FreeForm Modeling System for modifying. At last, auxiliary partial orthotopic liver transplantation was simulated with the force-feedback equipment (PHANTOM). Results ①It had greatly verisimilar image for the reconstructed 3D liver moulds with artery, vein, portal vein and bile duct; By liver seeing through, it had high fidelity and b 3D effect for the intrahepatic artery, vein, portal vein and bile duct, and their spatial disposition and course and correlationship were shown clearly. ②In the virtual surgery system, the virtual scalpel could be manipulated on 3D liver mould with PHANTOM. The simulating effect was the same as the clinic operation for auxiliary partial orthotopic liver transplantation. Plane visualization of hepatic resection and intrahepatic vessel cutting was achieved by adjusting the transparency of the resection part. Life-like could be felt and power feeling could be touched during virtul operation. Conclusion ①The visualized liver mould reconstructed is 3D and verisimilar, and it is helpful to design reasonable scheme for liver transplantation. ②It not only can improve the surgical effect and decrease the surgical risk, but also can reduce the complications and enhance the communication between doctor and patient through designing surgical plan and demonstrating visualized operation before surgery. ③Visualized liver transplantation surgery is helpful for medical workers to train and study.
ObjectiveTo explore value of multidisciplinary team (MDT) discussion in comprehensive downstaging treatment of liver cancer.MethodThe clinical data of 2 patients with liver cancer who could not undergo the radical surgery admitted to the Affiliated Hospital of Southwest Medical University were analyzed retrospectively.ResultsCase 1 was diagnosed as the liver cancer with extensive double lung metastasis at admission. The clinical stage was stage Ⅲb; After MDT discussion, the patient was treated with chemotherapy and embolization via hepatic artery and bronchial artery; At the same time, the patient was treated with apatinib; At present, the metastasis of both lungs disappeared completely; The clinical stage was stage ⅡB, and the radical resection was proposed. Case 2 was diagnosed as the right liver cancer at admission. The clinical stage was stage Ⅰ b. The preoperative examination showed that the hepatic reserve function was poor and the patient could not tolerate the half hepatectomy; After MDT discussion, the patient was treated with the combination of chemotherapy and embolization via the hepatic artery and apatinib in the same period; At the same time, the patient was treated with liver protection. The clinical stage was reduced to stage Ⅰ a. The hepatic reserve function improved and the laparoscopic right hemihepatectomy was performed, no recurrence or metastasis was found after 3 months follow-up.ConclusionComprehensive downstaging treatment based on MDT model could bring better clinical outcomes for patients with liver cancer who are unable to undergo one-stage radical surgery.
Objective To explore feasibility and safety of ex vivo liver resection and autotransplantation in treating end-stage hepatic alveolar echinococcosis combined with secondary cavernous transformation of portal vein. Methods The patient was diagnosed with the end-stage hepatic alveolar echinococcosis combined with secondary cavernous transformation of portal vein. The ultrasonography, computed tomography, and magnetic resonance imaging were used to access the characteristics of the lesions and the extent of involvement of the portal vein and its branches. The liver model was reconstructed using a three-dimensional imaging data analysis system (EDDA Technology, Inc. USA), the remnant liver volume and the extent of involvement of the first hepatic hilum were recorded. Then the multidisciplinary team repetitively discussed the risks and procedures involved in the surgery. Finally, the ex vivo liver resection and autotransplantation was proposed. Results The preoperative evaluation showed the patient had a large intrahepatic lesion which severely invaded the retrohepatic inferior vena cava, the right hepatic vein, and the middle hepatic vein and were completely occluded, the left hepatic vein was partially invaded, and the portal vein was spongiform. The remnant liver volume was 912 mL, the ratio of residual liver volume to standard liver volume was 0.81. The preoperative liver function Child-Pugh score was grade A. The ex vivo liver resection and autotransplantation was successfully managed according to the expected schedule. The autografts (made by patient’s great saphenous vein) were used to reconstruct the hepatic vein and portal vein, and the retrohepatic inferior vena cava was not reconstructed. The patient recovered well and was discharged on day 20 after the operation. Conclusions Ex vivo liver resection and autotransplantation could successfully be applied in treating patient with end-stage hepatic alveolar echinococcosis combined with secondary cavernous transformation of portal vein. Adequate preoperative assessment and management of the first hepatic hilum are key to this operation.
Objective The aim is to sort CD90+ subpopulation cells in human liver cancer cell lines and investigate efficiency of magnetic cell sorting (MACS) on sorting the liver cancer stem cells. Methods ①Expressions of CD90. Immunohistochemical method was used to determine the expressions of CD90 in normal liver tissues in 8 cases, liver cancer and adjacent liver cancer tissues in 58 cases. ②Screened the cell lines. Huh-7, MHCC97-H, Bel-7402, and SMMC-7721 cell lines were divided into blank control group and experimental group (5.5×105 cells per hole, 1 hole), cells of the experimental group were added with 5 μL CD90–PE while cells of the blank control group were treated with 5 μL CD90–PE non fluorescent antibody. Determined the proportion of CD90+ cells in the 2 groups by flow cytometry (FCM). ③MACS. Huh-7 and MHCC97-H cell lines were labeled with magnetic beads respectively and sorted by MACS, 1 mL cell suspensionsorted by magnetic sorting (MS) was collected as CD90– group, and 1 mL PBS after MS wash was collected as CD90+ group, as well as blank control group and experimental group. Determined the proportion of CD90+ cells in 4 groups by FCM. Two times of MACS were performed in Huh-7 cells. ④Serum free culture and serum culture. Huh-7 cells were divided into serum-free culture group and serum culture group (1 hole), and proportions of CD90+ cells were determined by FCM at 1 week after culture. Results ①The positive rate of CD90 was 0 (0/8), 65.5% (38/58), and 20.7% (12/58) in normal liver tissues, liver cancer tissues, and adjacent liver cancer tissues respectively, and the positive rate of CD90 was higher in liver cancer tissues than those of normal liver tissues (χ2=6.78, P<0.05) and adjacent liver cancer tissues (χ2=20.83, P<0.05). ②For Huh-7, MHCC97-H, SMMC-7721, and Bel7402 cell lines, the proportions of CD90+ cells in the experimental group was 0.851%, 1.090%, 2.710%, and 4.050% respectively, the proportions of CD90+ cells in the blank control group was 0.241%, 0.688%, 1.890%, and 2.080% respectively, so we chose Huh-7 and MHCC97-H cell lines to perform MACS. ③Results of MACS for Huh-7 cell line. For the first MACS, the proportions of CD90+ cells in the blank control group, experimental group, CD90– group, and CD90+ group was 0.241%, 0.851%, 0.574%, and 1.100% respectively. For the second MACS, the proportions of CD90+ cells in the blank control group, experimental group, CD90– group, and CD90+ group was 0.032%, 0.961%, 0.426%, and 9.700% respectively. Conclusions The normal liver tissues do not express the CD90, but the liver cancer tissues express CD90 highly. There is a few CD90+ cells in Huh-7 and MHCC97-H liver cancer cell lines. The MACS has a certain effect on improving the proportion of CD90+ cells in the cell lines. The serum-free suspension culture has no effect on enriching CD90+ cells.