PLCε基因敲除小鼠移植性肝癌模型的建立与分析

目的利用PLCε基因敲除小鼠建立移植性肝癌动物模型,以探讨PLCε基因在肝肿瘤生长发育过程中的作用。方法选取PLCε+/+小鼠15只为对照组,选取PLCε+/+(野生型)和PLCε-/-(敲基因型)小鼠各15只为模型组I和II,沿腹中线开腹后将H22细胞接种到模型组小鼠肝脏实质内,对照组注射生理盐水。于注射后第15天行剖腹探查,观察各组成瘤率及肿瘤体积,并进行肿瘤病理学分析。结果各组小鼠的存活率均为100%。模型组I肿瘤移植成功率为100%,肿瘤体积平均为(65.21±5.25)mm3。模型组II的肿瘤移植成功率为53.3%,肿瘤体积平均为(23.46±3.47)mm3。模型组I的肿瘤平均体积明显大于模型组II(P0.05)。模型I组和II组病理检查均证实为原位肝细胞瘤。结论成功建立了PLCε敲基因小鼠移植性肝癌动物模型,验证了PLCε敲基因小鼠具有抑制肝肿瘤生长的特性。     

Hepatic Phosphatidylethanolamine Methyltransferase Activity Is Decreased by Ethanol and Increased by Phosphatidylcholine

Phosphatidylethanolamine N-methyltransferase participates in the synthesis of membrane phosphatidylcholine. Its activity was reported to be decreased in patients with alcoholic cirrhosis, but it is not known whether this is a consequence of the cirrhosis or precedes it. This question was studied in a baboon model of alcohol-induced fibrosis. Phosphatidylethanolamine N-methyltransferase activity was measured in sequential percutaneous needle liver biopsies by the conversion of phosphatidylethanolamine to phosphatidylcholine, using radioactive S-adenosylmethionine as a methyl donor. Chronic alcohol consumption (1–6 years) significantly decreased hepatic phospholipid and phosphatidylcholine levels and reduced phosphatidyl-ethanolamine N-methyltransferase activity even before the development of fibrosis. These effects were prevented or attenuated by supplementing the diet with 2.8 g/1000 kcal of a preparation rich in dilinoleoyl phosphatidylcholine, a highly bioavailable phosphatidylcholine species. There were significant ( p < 0.001) correlations between phosphatidylethanolamine N-methyltransferase activity and both hepatic phosphatidylcholine ( r = 0.678) and total phospholipid ( r = 0.662).
Conclusions:

• 1. 

  Alcohol consumption diminishes phosphatidylethanolamine N-methyltransferase activity prior to the development of cirrhosis and decreases the hepatic content of its product, namely phosphatidylcholine, a key component of cell membranes. This may promote hepatic injury and possibly trigger fibrosis.
• 2. 

  Phosphatidylcholine administration ameliorates the ethanol-induced decrease in phosphatidylethanolamine N-methyltransferase activity and corrects phospholipid and phosphatidylcholine depletions, thereby possibly contributing to the protection against alcoholic liver injury.

     

Gastroduodenal artery disconnection during liver transplantation decreases non-anastomotic stricture incidence

BackgroundThe hepatic artery is the only blood source nourishing the biliary duct and associated with biliary complication after liver transplantation (LT). Gastroduodenal artery (GDA) disconnection increased proper hepatic artery flow. Whether this procedure attenuates biliary non-anastomotic stricture (NAS) is not clear.MethodsA total of 241 patients with LT were retrospectively analyzed. The patients were divided into the GDA disconnection (GDA-) and GDA preservation (GDA+) groups. Propensity score matching (PSM) was administrated to reduce bias. Logistic regression was conducted to analyze risk factors for biliary NAS before and after PSM. Postoperative complications were compared. Kaplan-Meier survival analysis and log-rank tests were performed to compare overall survival.ResultsIn all, 99 patients (41.1%) underwent GDA disconnection, and 49 (20.3%) developed NAS. Multivariate logistic regression revealed that GDA preservation (OR = 2.24, 95% CI: 1.11-4.53; P = 0.025) and model for end-stage liver disease (MELD) score > 15 (OR = 2.14, 95% CI: 1.12-4.11; P = 0.022) were risk factors for biliary NAS. PSM provided 66 pairs using 1:2 matching method, including 66 GDA disconnection and 99 GDA preservation patients. Multivariate logistic regression after PSM also showed that GDA preservation (OR = 3.15, 95% CI: 1.26-7.89; P = 0.014) and MELD score > 15 (OR = 2.41, 95% CI: 1.08-5.36; P = 0.031) were risk factors for NAS. When comparing complications between the two groups, GDA preservation was associated with a higher incidence of biliary NAS before and after PSM (P = 0.031 and 0.017, respectively). In contrast, other complications including early allograft dysfunction (P = 0.620), small-for-size graft syndrome (P = 0.441), abdominal hemorrhage (P = 1.000), major complications (Clavien-Dindo grade ≥ 3, P = 0.318), and overall survival (P = 0.088) were not significantly different between the two groups.ConclusionsGDA disconnection during LT ameliorates biliary NAS incidence and may be recommended for application in clinical practice.     

Sirolimus improves the prognosis of liver recipients with hepatocellular carcinoma: A single-center experience

BackgroundTumor recurrence after liver transplantation (LT) for selective patients diagnosed with hepatocellular carcinoma (HCC) in the setting of cirrhosis is the greatest challenge effecting the prognosis of these patients. The aim of this study was to evaluate the efficacy of sirolimus on the prognosis for these recipients.MethodsThe data from 193 consecutive HCC patients who had undergone LT from January 2015 to December 2019 were retrospectively analyzed. These patients were divided into the sirolimus group [patients took sirolimus combined with calcineurin inhibitors (CNIs) (n = 125)] and non-sirolimus group [patients took CNI-based therapy without sirolimus (n = 68)]. Recurrence-free survival (RFS) and overall survival (OS) were compared between the two groups. The prognostic factors and independent risk factors for RFS and OS were further evaluated.ResultsNon-sirolimus was an independent risk factor for RFS (HR = 2.990; 95% CI: 1.050-8.470; P = 0.040) and OS (HR = 3.100; 95% CI: 1.190-8.000; P = 0.020). A higher proportion of patients beyond Hangzhou criteria was divided into the sirolimus group (69.6% vs. 80.9%, P = 0.030). Compared with the non-sirolimus group, the sirolimus group had significantly better RFS (P < 0.001) and OS (P < 0.001). Further subgroup analysis showed similar results.ConclusionsThis study demonstrated that sirolimus significantly decreased HCC recurrence and prolonged RFS and OS in LT patients with different stage of HCC.     

共刺激分子B7-H3表达对肝细胞癌转移的影响及机制研究

目的 探讨共刺激分子B7-H3在肝细胞癌（HCC）转移中的作用及其机制。方法 选择术中或术后病理组织学证实存在转移的HCC患者37例（转移组）,以同期按1∶1匹配无转移患者（无转移组）为配对资料,采用免疫组化法检测HCC及其对应癌旁组织和转移灶中B7-H3的染色情况。设计针对B7-H3基因的shRNA沉默质粒转染肝癌HepG2细胞,采用RT-PCR和Western blotting分别检测转染前后上皮间质转化（EMT）机制相关分子E-钙粘蛋白、波形蛋白和N-钙粘蛋白mRNA和蛋白水平的变化。结果 原发灶边缘和转移灶较原发灶存在更高强度的B7-H3表达。转移组原发灶B7-H3染色强度明显高于无转移组（P=0.01）。B7-H3 shRNA转染组HepG2细胞中E-钙粘蛋白mRNA和蛋白表达分别为1.27±0.23和1.03±0.27,均高于对照质粒转染组（0.71±0.16,0.80±0.05）和未转染组（0.63±0.11,0.71±0.09）,差异有统计学意义（P＜0.05）。B7-H3 shRNA转染组波形蛋白mRNA和蛋白表达量为0.31±0.14和0.36±0.06,均低于对照质粒转染组（0.79±0.09,0.81±0.15）和未转染组（0.82±0.04,0.98±0.15）,差异有统计学意义（P＜0.05）。B7-H3 shRNA转染组N-钙粘蛋白mRNA和蛋白表达量为0.68±0.09和0.56±0.16,均低于对照质粒转染组（1.28±0.26,0.86±0.09）和未转染组（1.42±0.17,1.02±0.11）,差异有统计学意义（P＜0.05）。结论 共刺激分子B7-H3可通过调控EMT对HCC肿瘤转移发挥促进作用。     

Serum hepatitis B surface antigen correlates with fibrosis and necroinflammation: A multicentre perspective in China

The kinetics of serum hepatitis B surface antigen (HBsAg) during the natural history of hepatitis B virus (HBV) infection has been studied, but the factors affecting them remain unclear. We aimed to investigate the factors affecting HBsAg titres, using data from multicentre, large‐sized clinical trials in China. The baseline data of 1795 patients in 3 multicentre trials were studied, and the patients were classified into 3 groups: hepatitis B early antigen (HBeAg)‐positive chronic HBV infection (n = 588), HBeAg‐positive chronic hepatitis B (n = 596), and HBeAg‐negative chronic hepatitis B (n = 611). HBsAg titres in the different phases were compared, and multiple linear progression analyses were performed to investigate the implicated factors. HBsAg titres varied significantly in different phases (P = .000), with the highest (4.60 log10 IU/mL [10%‐90% confidence interval: 3.52 log10 IU/mL‐4.99 log10 IU/mL]) in patients with HBeAg‐positive chronic HBV infection. In all phases, age and HBV DNA were correlated with serum HBsAg level. In HBeAg‐positive chronic hepatitis B patients, a negative correlation between HBsAg titres and fibrosis stage was observed. Alanine amonitransferase or necroinflammatory activity was also correlated with HBsAg titres in HBeAg‐negative chronic hepatitis B patients. In conclusion, decreased HBsAg titres may be associated with advancing fibrosis in HBeAg‐positive chronic hepatitis B patients or increased necroinflammation in those with HBeAg‐negative chronic hepatitis B. Our findings may help clinicians better understand the kinetics of HBsAg and provide useful insights into the management of this disease.