目的 总结超声引导下经皮经肝穿刺胆管引流术(PTCD)的优、缺点,为临床治疗重症急性胆管炎（SAC）提供参考。方法 回顾性分析我院1994年8月至2008年7月期间对156例老年SAC患者行在超声引导下的PTCD治疗的临床资料。结果 156例行PTCD均获成功,1次穿刺成功140例，其成功率达89.7%（140/156）; 16例首次穿刺失败后再次穿刺均成功。无一例发生腹腔出血、胆汁性腹膜炎等并发症。本组引流效果较好,中毒危象缓解，黄疸减退，肝功能改善。结论 PTCD较外科手术创伤小、操作简单、快速,具有微创的特点,对老年、有严重合并症及复杂疾病不能耐受手术及麻醉的SAC患者，其作为紧急抢救措施切实可行,并为后期施行根治性手术争取了时间。
Objective To systematically review the effectiveness and model building process of heparin treatment for animal model with smoke inhalation injury. Methods Databases including PubMed, EMbase, CBM, CNKI, VIP and WanFang Data were searched to collect animal experiments about the treatment of heparin for animal model with smoke inhalation injury from inception to November 2016. Two reviewers independently screened literature, extracted data and assessed the risk of bias of included studies. Then meta-analysis was conducted by RevMan 5.3 software. Results A total of nine studies involving 11 animal experiments were included. The results showed that building animal model with smoke inhalation injury were through burning of cotton towels or pine sawdust by sheep or rats below 40℃. The results of meta-analysis showed that there was no significant difference in mortality rate between two groups (heparin group vs. control group: RR=0.38, 95%CI 0.14 to 1.05, P=0.06; heparin plus DMSO group vs. DMSO group: RR=0.10, 95%CI 0.01 to 1.51, P=0.10). In addition, the pulmonary artery pressure (MD=–3.31, 95%CI –4.51 to –2.11, P<0.000 01), wet to dry weight ratio (MD=–0.90, 95%CI –1.19 to –0.61, P<0.000 01), and lung water content (MD=–1.18, 95%CI –1.67 to –0.70, P<0.000 01) of the experimental group were lower than those in the control group. PaO2/FiO2 after 12 hours (MD=131.00, 95%CI 59.54 to 202.46, P=0.000 3), PaO2/FiO2 after 24 hours (MD=114.00, 95%CI 60.56 to 167.44, P<0.000 1), PaO2/FiO2 after 48 hours (MD=46.00, 95%CI 20.62 to 71.38, P=0.000 4) were higher than those in the control group. However, there was no significant difference in coagulation function between both groups. Conclusion The current evidence shows that the establishment of animal model of smoke inhalation injury is still lack of standard method. Heparin can decrease pulmonary artery pressure and lung water content in animal models with smoke inhalation injury. Due to the limited quality and quantity of included studies, the above conclusions are still needed to be verified by more high quality studies.
ObjectiveTo compare the effect and safety of basiliximab in ABO incompatible pediatric liver transplant recipients.MethodsABO incompatible pediatric liver transplantation operated between January 2019 and August 2020 were studied. The patients were allocated randomized into two groups. Patients in experimental group were treated with basiliximab as immune induction therapy, but basiliximab was not used in patients of control group. Tacrolimus combined methylprednisolone were used after liver transplantation. The clinical characteristics, graft and recipient survival rate, rejection, infectious complications, and kidney functions after liver transplantation were observed. Donor specific antibody (DSA) was tested in 3 months after liver transplantation. The growth and development were assessed too after liver transplant.ResultsFourty-four patients were enrolled in the study, including 19 patients in the experimental group and 25 patients in the control group. The median follow-up time was 16.6 months (3.8–25.4 months), and there were no statistically differences between the two groups in terms of age, sex, weight, pediatric end-stage liver disease (PELD) score, and other basic conditions. There were no significant differences between the two groups in tacrolimus dose, tacrolimus trough concentration, kidney functions, height and weight growth after liver transplantation. There were no statistical differences in lung infection, blood stream infection within 3 months after liver transplantation, cytomegalovirus, EBV infection, graft/patient survival rate after liver transplantation (P>0.05). However, the acute rejection rate was lower and the DSA positive rate in 3 months after liver transplantation was lower in the experimental group (P<0.05).ConclusionsBasiliximab can be safely used in ABO incompatible pediatric liver transplant recipients. Acute rejection rate and DSA positive rate after transplantation can be decreased with the useof basiliximab.
ObjectiveTo study the development of methods assessing donor liver viability in liver transplantation.MethodsThe literature in the recent years on the methods of assessing donor liver viability was reviewed.ResultsFrom donor liver morphology to function,there have being developed many methods which assess donor liver viability,including:①donor liver appearance; ②intraoperative biopsies; ③donor liver microcirculation; ④portal pressure; ⑤enzymes levels in liver; ⑥lidocainemetabolizing activity; ⑦energy metabolism of donor liver; ⑧fat content in donor liver.ConclusionThere are many methods to assess the viability of donor liver. Each has its supericrity and defect respectively. Intraoperative biopsies, 31Pmagnetic resonance spectroscopy and portal pressure have more importance in clinical application.
Objective To explore the effects of overexpression of human tissue inhibitors of metalloproteinase-1 (hTIMP-1) on proliferation of human liver cancer cell line HepG2 in vitro. Methods A recombinant adenoviral vector containing full-length cDNA of hTIMP-1 was generated and transfected into HepG2. The viral titer was checked by measuring GFP, and the expression of hTIMP-1 in vitro was detected by the techniques of Western blot and semi-quantitative RT-PCR. The ultrastructure was observed by transmission electron microscope and the effects of overexpression of hTIMP-1 on proliferation of HepG2 in vitro was analyzed by MTT assay and growth curve. Results The resultant AdhTIMP-1 was successfully constructed and the expression of hTIMP-1 was detected by Western blot and RT-PCR. The growth and proliferation of HepG2, which had been transfected with AdhTIMP-1, was significantly inhibited. Conclusion The proliferation of HepG2 was markedly inhibited by recombinant adenovirus-mediated overexpression of hTIMP-1, which may pave the way for further application in liver gene therapy.
ObjectiveTo explore risk factors of blood transfusion during liver transplantation and construct its prediction model. MethodsThe patients underwent liver transplantation who met the inclusion and exclusion criteria of this study from March 2020 to December 2020 in the Beijing Youan Hospital of Capital Medical University were retrospectively collected. The univariate and logistic multivariate analysis were used to evaluate the risk factors of blood transfusion during liver transplantation and construct the prediction model for intraoperative blood transfusion. ResultsA total of 151 eligible liver transplantation patients were collected in this study, including 51 non-transfusion patients and 100 transfusion patients. The univariate analysis results showed that the differences of primary diagnosis, preoperative hemoglobin (Hb), platelet count, prothrombin time, international normalized ratio, Child-Turcotte-Pugh score, and end-stage liver disease (MELD) score were statistically different between them (P<0.05). The above variables selected by the univariate analysis were selected by stepwise method, then the preoperative Hb and MELD score were selected into the multivariate logistic regression analysis, the results showed that the preoperative Hb≤113 g/L and MELD score ＞14 increased the risk of blood transfusion during liver transplantation [Hb: OR=6.652, 95%CI (2.282, 19.392), P<0.001; MELD score: OR=16.037, 95%CI (6.336, 40.592), P<0.001]. The logistic regression model predicted the area under receiver operating characteristic curve was 0.873 [95%CI (0.808, 0.919), P<0.001], the sensitivity and specificity were 91.0% and 67.5%, respectively, Youden index was 0.674, the accuracy was 86.1%. ConclusionsResults of this study suggest that preoperative Hb ≤113 g/L and MELD score＞14 increase risk of blood transfusion during liver transplantation. Logistic regression model constructed according to preoperative Hb and MELD score has a better sensitivity and specificity of intraoperative blood transfusion.
Primary human hepatocytes (PHH) are the gold standard of in vitro human liver model for drug screening. However, a problem of culturing PHH in vitro is the rapid decline of cytochrome P450 (CYP450) activity, which plays an important role in drug metabolism. In this study, thermo-responsive culture dishes were used to explore the conditions for murine embryonic 3T3-J2 fibroblasts to form cell sheet. Based on the cell sheet engineering technology, a three-dimensional (3D) “sandwich” co-culture system of 3T3-J2 cell sheet/PHH/collagen gel was constructed. The tissue structure and protein expression of the model section were observed by hematoxylin eosin staining and immunofluorescence staining respectively. Phenacetin and bupropion were used as substrates to determine the activity of CYP450. The contents of albumin and urea in the system were determined by enzyme linked immunosorbent assay (ELISA). The results showed that the complete 3T3-J2 cell sheet could be obtained when the cell seeding density was 1.5×106 /dish (35 mm dish) and the incubation time at low temperature was 60 min. Through cell sheet stacking, a 3D in vitro liver model was developed. Compared with the two-dimensional (2D) model, in the 3D model, the cell-cell and cell-matrix connections were tighter, the activities of cytochrome P450 CYP1A2 and cytochrome P450 CYP2B6 were significantly increased, and the secretion levels of albumin and urea were increased. These indexes could be maintained stably for 21 d. Therefore, cell sheet stacking is helpful to improve the level of liver function of 3D liver model. This model is expected to be used to predict the metabolism of low-clearance drugs in preclinical, which is of great significance for drug evaluation and other studies.
【 Abstract 】 Objective To probe into the role of inositol 1, 4, 5-trisphosphate (IP3) and bax gene expression in apoptosis of HepG2 cells induced by genistein (Gen). Methods HepG2 cells were treated with different concentrations including 20, 40, 60 and 80 μ mol/L Gen as HepG2 cells cultured with 0 μmol/L Gen for 72 h was control; HepG2 cells were treated with 60 μmol/L Gen for 6, 12, 24, 48 and 72 h as HepG2 cells treated with 60 μmol/L Gen for 0 h was control. IP3 content, bax mRNA expression and apoptosis rate were assayed by IP3- ［ 3H ］ Birtrak assay, RT-PCR and flow cytometry, respectively. ResultsHepG2 cells incubated with each concentration of Gen for 72 h ， IP3 content was lower than that of control 〔 (17.7 ± 1.3), (11.2 ± 0.9), (4.9 ± 0.5), (4.8 ± 0.3) pmol/106 cells vs (29.4 ± 0.5) pmol/106 cells 〕 , P ＜ 0.01 ； bax mRNA expression (RI which was the gray degree multiply area of bax/the gray degree multiply area of β -actin) was higher than that of control (0.26 ± 0.02, 0.33 ± 0.05, 0.35 ± 0.06, 0.38 ± 0.05 vs 0.09 ± 0.01), P ＜ 0.01 ； The apoptosis rate was higher than that of control 〔 (10.1 ± 0.9)%, (18.7 ± 1.6)%, (28.7 ± 2.5)%, (27.9 ± 2.0)% vs (2.6 ± 0.1)% 〕 , P ＜ 0.01. HepG2 cells were incubated with 60 μ mol/L Gen for 6, 12, 24, 48 and 72 h ， IP3 content was lower than that of control 〔 (22.6 ± 0.9), (12.0 ± 1.4), (7.5 ± 0.8), (5.6 ± 0.5), (4.3 ± 0.6) pmol/106 cells vs (29.2 ± 0.6) pmol/106 cells 〕 , P ＜ 0.01 ； bax mRNA expression was higher than that of control incubated with 60 μ mol/L Gen for above 12 h (0.25 ± 0.06, 0.29 ± 0.02, 0.30 ± 0.02, 0.35 ± 0.04 vs 0.09 ± 0.01), P ＜ 0.01 ； The apoptosis rate in groups incubated with 60 μ mol/L Gen for 24, 48 and 72 h was significantly higher than that in control 〔 (7.4 ± 0.5)%, (20.5 ± 2.0)%, (30.7 ± 1.6)% vs (2.6 ± 0.1)% 〕 , P ＜ 0.01. ConclusionGen induces apoptosis of HepG2 cells by reducing IP3 production and increasing bax gene expression.