Objectives To observe the expression of key proteins in the NLRP3/Caspase-1 pathway of pyroptosis in the mouse model of hepatic Echinococcus multilocularis (Em) infection and explore its correlation. Methods Twenty-five BALB/c mice were randomly divided into the control group and the infected group. The infected group was injected with 0.2 mL suspension of protoscolex (including 3 000 protoscoleces) injected under the liver capsule to establish a model of secondary infection with hepatic Em. The control group was treated without any treatments and conventional feeding was conducted. The mice were sacrificed at 1, 2, 3, and 5 months after infection. The liver was harvested and observed for gross morphology. HE staining and transmission electron microscopy were performed to observe the histopathological changes. The expressions of key proteins in the NLRP3/Caspase-1 pathway of pyroptosis and the IL-1β, a downstream factor of pyroptosis in the liver were detected by immunohistochemistry, Western blot and ELISA. Results Compared with the control group, the cystic lesions on the surface of liver tissues in the infected group mice gradually increased and protruded from the liver surface with the extension of infection time. HE staining showed various pathological changes such as inflammatory cell infiltration and fibrous hyperplasia in the liver lesions to varying degrees. After 2 months of Em infection, transmission electron microscope observation showed that the cell membrane of hepatocytes were broken and discontinuous, conforming to the "punching" phenomenon of pyroptosis. The results of ELISA showed that the concentration of IL-1β in liver homogenate of mice after 1, 2, 3 and 5 months of Em infection were significantly higher than that of the control group, and the difference was statistically significant (F=127.2, P<0.05). Immunohistochemical examination showed that the positive cell ratios of Caspase-1 and NLRP3 in liver of mice infected with Em at 1, 2, 3 and 5 months, were higher than that of the control group, and the difference were statistically significant (F=114.6, P<0.05; F=85.89, P<0.05). The Western blot results showed that the relative expression levels of Caspase-1, Xiaopi D, and NLRP3 proteins in the liver of infected mice showed a trend of first increasing (the expression of Caspase-1 and GSDMD reached their peak at 1 month of infection, while the expression of NLRP3 reached its peak at 2 months of infection) and then decreasing. There were statistically significant differences between the infection groups at different time points and the control group, as well as comparison between the infection groups at different time points there were also statistically significant differences (all P<0.05). Conclusion It is feasible to establish mouse Em infection model by “skin incision and liver puncture through abdominal muscle layer”. There is a new type of programmed cell death, pyroptosis, after Em infection in mouse liver. It may play a role in inflammation amplification through pyroptosis NLRP3/Caspase-1 pathway.
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 improve the prognosis of primary liver cancer with portal vein tumor thrombus （PVTT）， the progress of treatment for primary liver cancer with PVTT was reviewed. Methods The therapeutic approach and its efficacy for primary liver cancer with PVTT were summarized by literature search within recent years. Results PVTT is a common complication of primary liver cancer， the therapeutic approach are surgical resection， transcatheter arterial chemoembolization (TACE), intraportal venous therapy， radiotherapy， ablation therapy, molecular targeted therapy， etc. The excision rate for primary liver cancer with PVTT is low， the treatment is difficult and the outcome is dismal. It remains a poor prognosis at present， and the therapeutic effect need to be promoted. Conclusions The main treatment for primary liver cancer with PVTT should be surgical excision combine with other multidisciplinary comprehensive treatment to improve the survival in patients with PVTT， moreover， the therapeutic approach should be individualized and sequentially according to the patient’s condition and the type of PVTT.
Objective To study the effect of splenectomy on the anti-tumor immunity in rats with induced hepatocellular carcinoma (HCC). Methods At the second and fourth month of the induced HCC, the NK cell activity, TNF-α level and total lymphcyte in blood were measured in the group of splenectomy and the control group. Results There were no different in the total lymphcyte and TNF-α in the blood in two groups, but there were significant difference in the NK cell activity between the group of splenectomy and the control group (Ｐ＜0.05). Conclusion There are some change in the anti-tumor immunity after splenectomy in rats, in which NK cell activity is at low level continuously. TNF-α isn′t affected after the second month after splenectomy.
ObjectiveTo find out the risk factors affecting the prognoses and microvascular invasion (MVI) of patients with China Liver Cancer Staging-stageⅠ a (CNLC Ⅰ a) hepatocellular carcinoma (HCC). MethodsBased on the established inclusion and exclusion criteria, the clinicopathologic information and follow-up data of patients with CNLC Ⅰ a HCC were retrospectively collected, who underwent radical resection in the West China Hospital of Sichuan University from Jan. 2012 to Dec. 2016. The Cox proportional hazards regression was utilized to analyze the risk factors affecting the prognosis of patients with CNLC Ⅰ a HCC, and the non-conditional logistic regression was utilized to analyze the preoperative clinical indicators associating with MVI. ResultsA total of 300 patients with CNLC Ⅰ a HCC were included in this study, among which 51 (17.0%) cases accompanied with MVI. The follow-up period ranged from 2 to 104 months (median 39 months), with a recurrence time ranging from 2 to 104 months (median 52 months), and an overall survival time ranging from 3 to 104 months (median 98 months). During the follow-up period, postoperative recurrence occurred in 145 (48.3%) cases. The Cox proportional hazards regression analysis revealed that: tumor diameter >3 cm, presences of MVI and satellite nodules increased the risk of shortened recurrence time for the patients with CNLC Ⅰ a HCC (P<0.05); Factors including gamma-glutamyltranspeptidase level >60 U/L, tumor low differentiation, presences of MVI and satellite nodules were associated with shortened overall survival time for the patients with CNLC Ⅰ a HCC (P<0.05). The preoperative alpha-fetoprotein level ≥400 μg/L and tumor diameter >3 cm increased the risk of presence of MVI for the patients with CNLC Ⅰ a HCC [χ2=3.059, OR(95%CI)=2.357(1.047, 5.306), P=0.038; χ2=3.002, OR(95%CI)=2.301(1.026, 5.162), P=0.043]. ConclusionThe results of this study suggest that adopting corresponding strategies to address the risk factors affecting prognosis of patients with CNLC Ⅰ a HCC and the risk factors associated with MVI can have a significant clinical impact on improving surgical treatment outcomes for these patients.
Objective To examine the effect of zinc finger protein A20 on regeneration of small-for-sized liver allograft, graft rejection and recipient rat survival time. Methods Small-for-sized liver transplantation with 30% partial liver allograft was performed by using a b-rejection combination rat model of DA (RT1a) to Lewis (RT1l) rats. The rats were grouped into rAdEasy-A20 treatment group （A20 group）, the control empty Ad vector rAdEasy treatment group (rAdEasy group) and PS control treatment group (PS group). Ex vivo gene transfer in donor liver graft was performed through portal vein infusion. Animals were assessed for survival days, expression of A20 in liver graft, liver graft regeneration, hepatocyte apoptosis, graft rejection, NF-κB activation and ICAM-1 mRNA expression in liver graft sinusoidal endothelial cells (LSECs), number of liver graft infiltrating mononuclear cells (LIMCs) and the subproportion of NK/NKT cells, and serum IFN-γ level. Results Survival day of A20 group rats was prominently longer than that of PS group rats and rAdEasy group rats (P=0.001 8), whereas survival day of rAdEasy group rats was remarkably shorter than that of PS group rats (P=0.001 8). Regeneration of the small-for-sized liver allograft was markedly augmented by A20, BrdU labelling index of hepatocyte on postoperative day 4 was significantly increased in the A20 group compared with the PS group and rAdEasy group (P＜0.01). Hepatocyte apoptosis on postoperative day 4 was significantly inhibited by A20 (P＜0.01). On postoperative day 4, histologic examination revealed a mild rejection in the A20 group but a more severe rejection in the PS and rAdEasy groups. NF-κB activity and ICAM-1 mRNA expression in LSECs on postoperative day 1 were notably suppressed by A20 overexpression. Flow cytometry analysis showed a marked downregulation of LIMCs number by A20, including more prominent decrease in the subproportion of NK/NKT cells on postoperative day 1 and 4, respectively (P＜0.05). Serum IFN-γ level on postoperative day 4 was also significantly suppressed by A20 overexpression (P＜0.05). Conclusion These data suggest that A20 could effectively promote small-for-sized liver allograft regeneration, suppresses rejection and prolong survival days of recipient rats. These effects of A20 could be related to an inhibition of LSECs activation, suppression of infiltration of LIMCs and the subpopulations such as NK cells and NKT cells into liver graft, and inhibition of hepatocyte apoptosis.
ObjectiveTo study the clinical role of spleen/remnant liver volume ratio in evaluating liver reserve function after surgical treatment for liver cancer. MethodsTo calculate the ratio of spleen volume/remnant liver volume after tumor excision with imaging method and immersion method; to analyze the relationships between spleen/remnant liver volume ratio and liver function score after operation as well as hospital stay. ResultsLiver function ChildPugh score was related mainly with spleen/remnant liver volume ratio (t=7.831, P=0.000), which was proved by multiple regression analysis. The median hospital stay of the group with spleen/remnant liver ratio ≤0.9 was 14 d （12-16 d）, which was less than that (22 d, 15-29 d) of the other group with the ratio gt;0.9 (P=0.000). ConclusionsSpleen/remnant liver volume ratio can predict effectively recovery ability of patients after operation for liver cancer， and assess correctly the reserve function of liver. When the ratio is less than or equal 0.9, the operation is safe.
ObjectiveTo evaluate the safety and technical feasibility of salvage liver transplantation (SLT) after liver resection, and its influence on prognosis. MethodsThe clinical data of 289 patients who underwent liver transplantation by cadaveric grafts treating for hepatocellular carcinoma met the UCSF criteria from June 2001 to December 2008 were analyzed retrospectively. Among them, 242 patients underwent primary liver transplantation (PLT, PLT group), and 47 patients underwent SLT for recurrence (SLT group). Perioperative factors and survival were compared between two groups. ResultsThere were no significant differences of age, gender, and pathology of tumor between two groups (Pgt;0.05). The operation time in the SLT group was significantly longer than that in the PLT group 〔(7.1±1.8) h versus (6.4±1.4) h， P=0.004〕. The differences of intraoperative blood loss 〔(2 560.5±2 683.6) ml versus (2 042.9±2 006.2) ml， P=0.173〕 and blood transfusion 〔(13.8±12.9) U versus (9.9±12.6) U， P=0.087〕 were not significant between two groups. The mean interval time from resection to transplantation was (32.8±32.4) months. The median followup was 38.7 months, 3year overall and diseasefree survivals were not significantly different (82.3% versus 75.5%, P=0.312; 78.8% versus 70.1%, P=0.755, respectively) between the SLT group and PLT group. According to intentiontotreat analysis, the 3year overall survival in the SLT group was significantly longer than that in the PLT group (88.4% versus 76.2%, P=0.047). ConclusionsIn selected patients, liver resection prior to transplantation neither increases operative morbidity nor impairs prognosis following liver transplantation. SLT after liver resection, can be an alterative treatment for HCC.
In order to solve the pathological grading of hepatocellular carcinomas (HCC) which depends on biopsy or surgical pathology invasively, a quantitative analysis method based on radiomics signature was proposed for pathological grading of HCC in non-contrast magnetic resonance imaging (MRI) images. The MRI images were integrated to predict clinical outcomes using 328 radiomics features, quantifying tumour image intensity, shape and text, which are extracted from lesion by manual segmentation. Least absolute shrinkage and selection operator (LASSO) were used to select the most-predictive radiomics features for the pathological grading. A radiomics signature, a clinical model, and a combined model were built. The association between the radiomics signature and HCC grading was explored. This quantitative analysis method was validated in 170 consecutive patients (training dataset: n = 125; validation dataset, n = 45), and cross-validation with receiver operating characteristic (ROC) analysis was performed and the area under the ROC curve (AUC) was employed as the prediction metric. Through the proposed method, AUC was 0.909 in training dataset and 0.800 in validation dataset, respectively. Overall, the prediction performances by radiomics features showed statistically significant correlations with pathological grading. The results showed that radiomics signature was developed to be a significant predictor for HCC pathological grading, which may serve as a noninvasive complementary tool for clinical doctors in determining the prognosis and therapeutic strategy for HCC.
ObjectiveTo summarize experiences of surgical treatment of complex giant cavernous hemangioma of the liver. MethodThe clinical data of 55 patients with complex hepatic cavernous hemangioma with tumor diameter more than 10 cm and in close proximity to hepatic hilar region or vena cava inferior underwent surgical treatment from January 2009 to December 2014 were analyzed retrospectively. ResultsAmong these 55 patients with complex giant cavernous hemangioma,13 cases (23.6%) were male,42 cases (76.4%) were female.The median age was 49.2 years (range from 23 to 68 years).Hepatic hemangioma with multiple lesions was most common (71.0%,39/55).The tumor happened mostly in the right hepatic lobe (47.3%,26/55).The median size of complex giant cavernous hemangioma was 16.2 cm (10.2-50.0 cm).The liver functions of all the patients were normal (Child-Pugh A).Different methods of hepatic inflow occlusion and surgical procedures were performed according to the tumor location and size.Of the patients,17 cases were underwent Pringle maneuver,12 cases were underwent modified Pringle maneuver and 1 case was underwent hemihepatic vascular occlusion;28 cases were treated by extracapsular enucleation,27 cases by liver resection.The average operative time was 202 min (85-420 min).The average intraoperative blood loss was 855.5 mL (50-3 000 mL).Twenty-six cases (47.3%) had no blood transfusion,and 10 cases (18.2%) had autologous blood transfusion.The associated complications occurred in 7 patients after surgery,and no surgical death occurred.The median postoperative hospital stay was 14.8 d. ConclusionsThe essential points in operation for the complex giant cavernous hemangioma are the control and management of the operative massive bleeding,and the preservation of the normal hepatic parenchyma as much as possible.The surgical treatment is safe and feasible under the proper hepatic inflow occlusion and resection methods.The prevention and management of bile leakage is also important.