Pretransplant use of immune checkpoint inhibitors for hepatocellular carcinoma: A multicenter, retrospective cohort study.

Immune checkpoint inhibitors (ICIs) as a downstaging or bridging therapy for liver transplantation (LT) in hepatocellular carcinoma patients are rapidly increasing. However, the evidence about the feasibility and safety of pre-LT ICI therapy is limited and controversial. To this end, a multicenter, retrospective cohort study was conducted in 11 Chinese centers. The results showed that 83 recipients received pre-LT ICI therapy during the study period. The median post-LT follow-up was 8.1 (interquartile range 3.3-14.6) months. During the short follow-up, 23 (27.7%) recipients developed allograft rejection, and 7 of them (30.4%) were diagnosed by liver biopsy. Multivariate logistics regression analysis showed that the time interval between the last administration of ICI therapy and LT (TLAT) ≥ 30 days was an independent protective factor for allograft rejection (odds ratio = 0.096, 95% confidence interval 0.026-0.357; P < .001). Multivariate Cox analysis showed that allograft rejection was an independent risk factor for overall survival (hazard ratio = 9.960, 95% confidence interval 1.006-98.610; P = .043). We conclude that patients who receive a pre-LT ICI therapy with a TLAT shorter than 30 days have a much higher risk of allograft rejection than those with a TLAT longer than 30 days. The presence of rejection episodes might be associated with higher post-LT mortality.

Cancer-associated fibroblasts induce sorafenib resistance of hepatocellular carcinoma cells through CXCL12/FOLR1.

BACKGROUND: Due to the high drug resistance of hepatocellular carcinoma (HCC), sorafenib has limited efficacy in the treatment of advanced HCC. Cancer-associated fibroblasts (CAFs) play an important regulatory role in the induction of chemoresistance. This study aimed to clarify the mechanism underlying CAF-mediated resistance to sorafenib in HCC. METHODS: Immunohistochemistry and immunofluorescence showed that the activation of CAFs was enhanced in HCC tissues. CAFs and paracancerous normal fibroblasts (NFs) were isolated from the cancer and paracancerous tissues of HCC, respectively. Cell cloning assays, ELISAs, and flow cytometry were used to detect whether CAFs induced sorafenib resistance in HCC cells via CXCL12. Western blotting and qPCR showed that CXCL12 induces sorafenib resistance in HCC cells by upregulating FOLR1. We investigated whether FOLR1 was the target molecule of CAFs regulating sorafenib resistance in HCC cells by querying gene expression data for human HCC specimens from the GEO database. RESULTS: High levels of activated CAFs were present in HCC tissues but not in paracancerous tissues. CAFs decreased the sensitivity of HCC cells to sorafenib. We found that CAFs secrete CXCL12, which upregulates FOLR1 in HCC cells to induce sorafenib resistance. CONCLUSIONS: CAFs induce sorafenib resistance in HCC cells through CXCL12/FOLR1.

Follistatin Alleviates Hepatic Steatosis in NAFLD via the mTOR Dependent Pathway.

Purpose: In this study, we aimed to investigate the effect of follistatin (FST) on hepatic steatosis in NAFLD and the underlying mechanism, which has rarely been reported before. Methods: Liver samples from NAFLD patients and normal liver samples (from liver donors) were collected to investigate hepatic FST expression in humans. Additionally, human liver cells (LO2) were treated with free fatty acid (FFA) to induce lipid accumulation. Furthermore, lentivirus with FST overexpression or knockdown vectors were used to generate stable cell lines, which were subsequently treated with FFA or FFA and rapamycin. In the animal experiments, male C57BL/6J mice were fed with a high-fat diet (HFD) to induce NAFLD, after which the adeno-associated virus (AAV) gene vectors for FST overexpression were administered. In both cell culture and mice, we evaluated morphological changes and the protein expression of sterol regulatory element-binding protein1 (SREBP1), acetyl-CoA carboxylase1 (ACC1), carbohydrate-responsive element-binding protein (ChREBP), fatty acid synthase (FASN), and Akt/mTOR signaling. The body weight and serum parameters of the mice were also measured. Results: Hepatic FST expression level was higher in NAFLD patients compared to normal samples. In LO2 cells, FST overexpression alleviated lipid accumulation and lipogenesis, whereas FST knockdown aggravated hepatic steatosis. FST could regulate Akt/mTOR signaling, and the mTOR inhibitor rapamycin abolished the effect of FST knockdown on hepatic de novo lipogenesis (DNL). Furthermore, FST expression was increased in HFD mice compared to the corresponding controls. FST overexpression in mice reduced body weight gain, hyperlipidemia, hepatic DNL, and suppressed Akt/mTOR signaling. Conclusion: Hepatic FST expression increases in NAFLD and plays a protective role in hepatic steatosis. FST overexpression gene therapy alleviates hepatic steatosis via the mTOR pathway.Therefore, gene therapy for FST is a promising treatment in NAFLD.

Efficacy of Adjuvant Transarterial Chemoembolization after Radical Hepatectomy in Solitary Hepatocellular Carcinoma Patients: A Retrospective Study.

Background: More and more studies have suggested that hepatocellular carcinoma (HCC) patients with high-risk recurrence factors can benefit the most from postoperative adjuvant transarterial chemoembolization (PA-TACE) for its potential effect in delaying cancer recurrence. However, it remains unclear if solitary HCC (SHCC) patients particularly those without high-risk recurrence factors should also receive PA-TACE. This study aimed to analyze the efficacy of PA-TACE in them. Methods: Retrospectively, we enrolled 123 SHCC patients who either received radical hepatectomy alone (No TACE group, n = 39) or followed by PA-TACE (PA-TACE group, n = 84) in our institution. Prognostic risk factors, disease-free survival (DFS), and overall survival (OS) were analyzed using the Cox proportional hazard regression model, the Kaplan-Meier method, and the log-rank test. Results: Liver cirrhosis was the only independent risk factor for SHCC patients. Overall, the PA-TACE group had no improved OS (P = 0.977) but worse DFS compared with the No TACE group (P = 0.045). Consistently, in subgroup analysis, SHCC patients with negative microvascular invasion (MVI), tumor size ≤ 5 cm and preoperative alpha-fetoprotein (AFP) < 400 ng/ml had similar OS (P = 0.466, P = 0.864, P = 0.488, respectively) but even worse DFS (P = 0.035, P = 0.040, P = 0.019, respectively) than those in the No TACE group. Besides, there was no significant difference in DFS and OS between the two groups of SHCC patients with liver cirrhosis (P = 0.342, P = 0.941, respectively). Conclusions: PA-TACE may not improve the long-term survival of SHCC patients, but may even potentially promote their postoperative tumor recurrence, especially for those with MVI-negative, tumor size ≤ 5 cm, and preoperative AFP < 400 ng/ml.

Evidence of SARS-CoV-2 infection in gallbladder and aggravating cholecystitis to septic shock: a case report.

Coronavirus disease 2019 (COVID-19) has threatened human health worldwide and could lead to multiple organs injury. However, the impact on the virus infecting the biliary system, especially the gallbladder, has remained unclear and no pathological evidence has been reported yet. A case of SARS-CoV-2 infection in a gallbladder with cholecystitis, which progressed rapidly to sepsis and required an emergency operation was investigated and reported. Clinical specimens of the COVID-19 patient including serum, oropharyngeal swabs, sputum, bile, abdominal drainage fluid, urine, stool, and gallbladder tissue were collected and tested for SARS-CoV-2 RNA using a quantitative polymerase chain reaction (qPCR) assay. Fresh normal gallbladder tissue and gangrenous gallbladder tissue were also collected for further research including hematoxylin and eosin (HE), immunohistochemistry (IHC), and immunofluorescent (IF) staining, and compared with the gallbladder from the COVID-19 patient. The bile, as well as the serum, oropharyngeal swabs, sputum, abdominal drainage fluid, urine, and rectal swabs were consecutively negative for SARS-CoV-2 RNA. The viral host receptor angiotensin-converting enzyme 2 (ACE2) was highly expressed in gallbladder epithelial cells, and viral nucleocapsid protein (NP) was visualized in the cytoplasm of gallbladder epithelial cells. Immune cells including CD2, CD3, CD4, CD8, CD20, CD38, CD68, and MPO were positive in gangrenous gallbladder tissues without SARS-CoV-2 infection, and were relatively downregulated in SARS-CoV-2 infective gallbladder tissue. This study provided evidence of SARS-CoV-2 infection in the gallbladder and verified that the gallbladder was one of the target organs that SARS-CoV-2 could attack and damage using ACE2 as a cell receptor. Due to the immune dysregulation involved, more vigilant management and early assessment is needed for COVID-19 patients with the comorbidity of cholecystitis.

SSR2 overexpression associates with tumorigenesis and metastasis of Hepatocellular Carcinoma through modulating EMT.

Background: Hepatocellular carcinoma (HCC) is a common malignancy around the world. The molecular mechanisms underlying HCC tumorigenesis and metastasis are far from clear. Numerous studies have pointed out that signal sequence receptor (SSR) is an endoplasmic reticulum-related protein involved in protein folding and processing of eukaryotic cells. SSR2 is a subunit of SSR protein, but the role of SSR2 in hepatocellular carcinoma is largely unknown and warrants further study. Materials and Methods: Several public databases were data mined to analyze the expression of four subunits of SSR between tumor and its peritumor counterparts. Also, the expression of SSR2 in our own collected tissues from HCC patients were analyzed by IHC and quantitative PCR. Survival analyses were conducted to delineate the prognostic value of SSR2. Clinical data were obtained followed by analysis based on SSR2 expression. Afterwards, cell proliferation, migration and invasion were detected by IncuCyte and trans-well assays, respectively. RNA interference was carried out by transfecting specific siRNA targeting SSR2 into cells using lipo2000. Western blot was applied to validate the knockdown effect and regulation on EMT-related proteins. Results: We examined the expression of SSR and its correlation with recurrence and survival of patients. We discovered that SSR2 overexpression was negatively associated with survival of HCC patients from TCGA databases and the mutation of SSR2 was most among the four subunits of SSR protein. Additionally, in this study, we collected tumor and adjacent tissues from 125 cases of HCC patients. Through constructing tissue microarray, we have identified that SSR2 was highly expressed in HCC tumor tissues compared with adjacent normal tissues of hepatocellular carcinoma patients by immunohistochemistry assays. Furthermore, Kaplan-Meier survival analysis from our collected tissues revealed that the overexpression of SSR2 was inversely correlated with disease free survival and overall survival of HCC patients. We elucidated that SSR2 promotes proliferation, migration and invasion of HCC cells. SSR2 knockdown suppressed epithelial mesenchymal transition (EMT) of HCC cells. Conclusions: These results collectively show that SSR2 is overexpressed in HCC tumor tissues, and it is an important factor in predicting survival of HCC patients. Additionally, it is involved in metastasis of HCC. These findings may help to exploit SSR2 as a novel factor in predicting prognosis and metastasis of HCC.

Polysulfone foam with high expansion ratio prepared by supercritical carbon dioxide assisted molding foaming method.

Polysulfone (PSU) is considered as an important candidate for the fabrication of high-performance microcellular polymers, but the preparation of PSU foam with a high expansion ratio still remains a big challenge worldwide. In this study, high expansion ratio PSU foam was successfully prepared by a supercritical carbon dioxide (CO2) assisted molding foaming method. The foaming behavior of PSU under supercritical CO2 was systematically studied in various process conditions and different microcellular structures were created in PSU foams. The results showed that foaming temperature and CO2 concentration were the key factors to obtain microcellular foams with tailored microstructures. The cellular structure and expansion ratio of PSU foam obviously changed with different foaming temperatures. The expansion ratio and average cell size firstly increased and then decreased as foaming temperature increased. However, the cell density decreased and then remained stable as foaming temperature increased. The maximum expansion ratio of 11.0 was reached at the optimum foaming temperature of 200 °C. Cellular structure and morphologies of the foam changed obviously at CO2 concentrations below 5% and remained stable at CO2 concentrations above 5%. Finally, the prepared PSU foams exhibit excellent mechanical strength, good thermal conductivity, and superb heat retardancy, thus may have great potential application as a kind of substitute material in the electrical wire and cable industry, railway and steamer transportation, oil and gas platforms, military use and in other fields.

[Predicting value of dynamic alteration of blood neutrophil/lymphocyte ratio on recurrence-free survival in patients with advanced colon cancer after operation and chemotherapy].

OBJECTIVE: To evaluate whether neutrophil-lymphocyte ratio(NLR) predicts risk of recurrence in patients with advanced colon cancer undergoing curative resection followed by adjuvant chemotherapy. METHODS: A total of 149 patients with advanced colon cancer undergoing curative resection followed by adjuvant chemotherapy(FOLFOX6 protocol) were included. NLR was calculated preoperatively and before chemotherapy. The changes in NLR and the predictive value of NLR for prognosis were analyzed. RESULTS: The NLR of 149 patients was 2.8±1.5. NLR of 3.5 was identified according to the ROC curve. NLR<3.5 and NLR≥3.5 were classified as low and high NLR group, respectively. The 5-year recurrence-free survival(RFS) of patients with high preoperative NLR(n=22) was significantly worse than that of those with low preoperative NLR(n=127)(50.9% vs. 76.4%, P=0.025). The difference of 5-year RFS between high pre-chemotherapy NLR group(n=34) and low pre-chemotherapy NLR group(n=115) was statistically significant(50.1% vs. 71.4%, P=0.032). The 5-year RFS was 79.5% in patients with low preoperative NLR converting to high pre-chemotherapy NLR(n=16), similar to the group with high pre-chemotherapy group(P=0.077). The 5-year RFS was 17.7% in patients with high preoperative NLR reverting to low pre-chemotherapy NLR(n=12), similar to the group with low pre-chemotherapy group(P=0.978). There was significant difference in 5-year RFS between the postoperatively elevated group and postoperatively decreased group(P=0.036). CONCLUSION: An elevated blood NLR may be a biomarker of poor RFS in patients with advanced colon cancer after curative resection and chemotherapy.

[Midterm results of thoracic aortic dissection endovascular repair in conjunctions with the location of Adamkiewicz artery].

OBJECTIVE: To evaluate the effects of using longer xenografts in conjunctions with the location of Adamkiewicz artery (AKA) on midterm outcomes of endovascular treatment for thoracic aortic dissection. METHODS: From March 2005 to September 2011, 217 patients with type B dissection were recruited. There were 143 males and 74 females with a mean age of 65 ± 11 years. Among them, 43 patients were from Fifth Affiliated Hospital of Sun Yat-Sen University while another 174 patients from Affiliated Zhongshan Hospital of Fudan University. They were divided into 2 groups according to whether AKA was identified or not pre-operatively. Endovascular repairs were performed for all patients. Distal landing levels of xenografts were recorded. The thrombosis of false lumen and the complications of spinal cord injury and endoleak were analyzed. RESULTS: AKA was detected in 121 (55.8%) patients (group A) but not in 96 (44.2%) patients (group B). According to the levels of AKA, the patients of group A obtained the stabilization of affected thoracic aorta over a longer distance. And the ratio of patients with distal landing levels at T8-T10 was significantly higher than in group B (59.5% vs 12.5%, χ² = 49.85, P < 0.01). Also, during the follow-up period of 7.3 months, the ratio of patients with total thrombosis of false lumen in group A was significantly higher than that in group B (32.1% vs 19.1%, χ² = 4.34, P < 0.05). CONCLUSION: During the endovascular repair of thoracic aortic dissection, selecting a longer device may provide a better structural stability of affected aorta and promote false lumen thrombosis.

[Effect of suprarenal fixation on renal function in endovascular abdominal aneurysm repair patients].

OBJECTIVE: to assess the the mid-term renal function of abdominal aortic aneurysm (AAA) patients following supra-renal endovascular repair. METHODS: from March 2005 to December 2009, 290 AAA patients were included and grouped depending upon whether they had received infra-renal (IR) or supra-renal (SR) EVAR. SR was performed in 173 patients, with a mean age of (72 ± 8) years and 85.0% for male. IR was performed in 117 patients, with a mean age of (71 ± 9) years and 90.6% for male. Preoperative and 1 week, 1-, 3-, 6-, 12-month postoperative serum creatinine (Cr) and cystatin C (Cys-C) were detected. Estimated glomerular filtration rate (eGFR) were calculated by Cystatin-based formula and Cr-based Cockcroft formula. T test were used to determine statistical difference between or within groups. RESULTS: all Patients received Talent or Zenith endograft. The ratio of device-used were 67/106 in SR group and 25/92 in IR group (P < 0.05). The other characteristics and operative files in two groups were well matched. Preoperative Cr and Cys-C were (82 ± 8) µmol/L and (0.89 ± 0.11) mg/L for SR group, (81 ± 11) µmol/L and (0.87 ± 0.15) mg/L for IR group, no difference between groups. Compared to preoperative renal markers within each group, Cr, Cys-C and eGFR worsening were found at 1 week and 12 months postoperative (P < 0.05). At 1 week postoperative, Cr in SR group and IR group were (98 ± 11) µmol/L and (95 ± 13) µmol/L, Cys-C were (1.01 ± 0.10) mg/L or (0.99 ± 0.10) mg/L. At 12 months postoperative, Cr in SR group and IR group were (91 ± 15) µmol/L or (90 ± 12) µmol/L, Cys-C were (1.03 ± 0.20) or (1.02 ± 0.21) mg/L. Also, Cys-C [SR: (0.93 ± 0.17) mg/L, IR: (0.92 ± 0.31) mg/L] and eGFR by Cys-C worsening were found at 6 months postoperative. There was no difference between groups in patients Cr, Cys-C and eGFR at each follow-up time interval. CONCLUSION: the use of SR fixation was not significantly associated with mid-term postoperative renal injury.

[Establishment of rabbit model of renal allograft transplantation using microsurgical technique].

OBJECTIVE: To establish rabbit model of renal allograft transplantation with reduced complications and high survival rate using microsurgical technique. METHODS: Twelve healthy adult rabbits were randomly divided into 2 groups of equal number, one as donor group and the other recipient. The left kidneys of the donor rabbits were removed followed by immediate reperfusion with 4 degrees celsius H-CA solution, before they were transplanted into the recipient rabbits with their left kidneys excised and end-to-end anastomosis of the renal arteries, veins and ureter respectively performed with microsurgical technique. Another 12 normal rabbits received operations to temporarily block the right renal arteries and veins, serving as control group, in which 11 completed the experiment. RESULTS: No thrombosis or stricture occurred at the site of anastomosis in rabbits with renal allograft transplantation, and the survival rate reached 91.7% (11/12). CONCLUSION: This rabbit model of renal allograft transplantation has markedly fewer complications with improved survival rate, thus providing a more practical and reliable model for experimental and clinical studies of renal transplantation.

Management of acute rejection of kidney allograft.

OBJECTIVE: To evaluate the management of acute rejection (AR) after kidney transplantation and investigate the factors influencing the clinical outcome of the patients. METHODS: A retrospective study was conducted in 86 cases of AR developed after primary kidney transplantation in the light of therapeutic measures, clinical outcome and prognosis. RESULTS: Among these patients, 81 survived AR after treatment. In patients with pulse treatment with methylprednisolone (MP), 48 out of 68 managed to survive the crises, while in those who received ATG as the first line drug therapy 10 out of 11 patients survived and in other cases, 6 out of 7 did due to first-line OKT3 administration. All the 20 patients who did not respond to MP received ATG or OKT3 instead, with 14 recovered. Of the 8 patients who failed to be cured by the management above, 6 with previous CSA treatment took FK506 and 3 were consequently cured. Five patients lost the allografts because of uncontrollable infection, allograft rupture or thrombosis. CONCLUSIONS: MP therapy is still the most commonly used primary treatment for acute rejection episodes. Increase of SCr by more than 10% on days 2 and 3 of MP therapy indicates poor prognosis. ATG or OKT3 can be effective against acute rejection not only as first-line but also as second-line drug. In condition of steroid-resistant rejection when ATG and OKT3 fail to manage, a change to baseline immunosuppression may be considered as the replacement of CSA with FK506.