Calcyclin-binding protein contributes to cholangiocarcinoma progression by inhibiting ubiquitination of MCM2

Background:Cholangiocarcinoma(CCA)represents the epithelial cell cancer with high aggressiveness whose five-year survival rate is poor with standard treatment.Calcyclin-binding protein(CACYBP)shows aberrant expression within several malignant tumors,but the role of CACYBP in CCA remains unknown.Methods:Immunohistochemical(IHC)analysis was used to identify CACYBP overexpression in clinical samples of CCA patients.Moreover,its correlation with clinical outcome was revealed.Furthermore,CACYBP’s effect on CCA cell growth and invasion was investigated in vitro and in vivo using loss-of-function experiments.Results:CACYBP showed up-regulation in CCA,which predicts the dismal prognostic outcome.CACYBP had an important effect on in-vitro and in-vivo cancer cell proliferation and migration.Additionally,knockdown of CACYBP weakened protein stability by promoting ubiquitination of MCM2.Accordingly,MCM2 up-regulation partly reversed CACYBP deficiency’s inhibition against cancer cell viability and invasion.Thus,MCM2 might drive CCA development by Wnt/β-catenin pathway.Conclusions:CACYBP exerted a tumor-promoting role in CCA by suppressing ubiquitination of MCM2 and activating Wnt/β-catenin pathway,hence revealing that it may be the possible therapeutic target for CCA treatment.

Appropriate treatment strategies improve survival of hepatocellular carcinoma patients with portal vein tumor thrombus

AIM: To evaluate the survival benefits of different treatment strategies for hepatocellular carcinoma (HCC) patients with portal vein tumor thrombus (PVTT) and to determine the prognosis factors.

Expert consensus on management of metabolic disease in Chinese liver transplant recipients

Metabolic disease,including diabetes mellitus,hypertension,dyslipidemia,obesity,and hyperuricemia,is a common complication after liver transplantation and a risk factor for cardiovascular disease and death.The development of metabolic disease is closely related to the side effects of immunosuppressants.Therefore,optimization of the immunosuppressive regimen is very important for the prevention and treatment of metabolic disease.The Chinese Society of Organ Transplantation has developed an expert consensus on the management of metabolic diseases in Chinese liver transplant recipients based on recent studies.Emphasis is placed on the risk factors of metabolic diseases,the effect of immunosuppressants on metabolic disease,and the prevention and treatment of metabolic diseases.

Effects of α-adrenoreceptor antagonists on apoptosis and proliferation of pancreatic cancer cells in vitro

AIM: To discuss the expression of α-adrenoreceptors in pancreatic cancer cell lines PC-2 and PC-3 and the effects of α1- and α2-adrenoreceptor antagonists, yohimbine and urapidil hydrochloride, on the cell lines in vitro. METHODS: We cultured the human ductal pancreatic adenocarcinoma cell lines PC-2 and PC-3 and analyzed the mRNA expression of α1- and α2-adrenergic receptors by reverse transcription polymerase chain reaction (RT-PCR). The effects of yohimbine and urapidil hydrochloride on cell proliferation were assessed by 3-(4,5-dimethylthiasol-2-yl)- 2,4,-diphenyltetrazolium bromide (MTT) assay. Apoptosis was detected using the terminal deoxyribonucleotidyl transferase (TdT)-mediated biotin-16-dUTP nick-end labeling (TUNEL) assay and flow cytometry (FCM). RESULTS: PC-2 expressed mRNA in α1- and α2- adrenoreceptors. MTT assays showed that urapidil hydrochloride had no effect on PC-3 cell lines. However, exposure to urapidil hydrochloride increased DNA synthesis in PC-2 cell lines as compared to the control group. PC-2 cell lines were sensitive to both drugs. The proliferation of the 2 cell lines was inhibited by yohimbine. Cell proliferation was inhibited by yohimbine via apoptosis induction. CONCLUSION: The expression of α1- and α2- adrenoreceptors is different in PC-2 and PC-3 cell lines, which might be indicative of their different functions. The α2-adrenoceptor antagonist, yohimbine, can inhibit theproliferation of both cell lines and induce their apoptosis, suggesting that yohimbine can be used as an anticancer drug for apoptosis of PC-2 and PC-3 cells.

Effect of glial cells on remyelination after spinal cord injury

Remyelination plays a key role in functional recovery of axons after spinal cord injury.Glial cells are the most abundant cells in the central nervous system.When spinal cord injury occurs,many glial cells at the lesion site are immediately activated,and different cells differentially affect inflammatory reactions after injury.In this review,we aim to discuss the core role of oligodendrocyte precursor cells and crosstalk with the rest of glia and their subcategories in the remyelination process.Activated astrocytes influence proliferation,differentiation,and maturation of oligodendrocyte precursor cells,while activated microglia alter remyelination by regulating the inflammatory reaction after spinal cord injury.Understanding the interaction between oligodendrocyte precursor cells and the rest of glia is necessary when designing a therapeutic plan of remyelination after spinal cord injury.

Chimeric antigen receptor-engineered T-cell therapy for liver cancer

Background: Chimeric antigen receptor-engineered T-cell(CAR-T) therapy is a newly developed immunotherapy used in the treatment of cancers. Because CAR-T therapy has shown great success in treating CD19-positive hematological malignancies, its application has been explored in the treatment of solid tumors, such as liver cancer. In this review, we discuss the immune characteristics of liver cancer, the obstacles encountered during the application of CAR-T therapy, and preclinical and clinical progress in the use of CAR-T therapy in patients with liver cancer.Data sources: The data on CAR-T therapy related to liver cancers were collected by searching Pub Med and the Web of Science databases prior to December 2017 with the keywords "chimeric antigen receptor","CAR-T", "liver cancer", "hepatocellular carcinoma", and "solid tumor". Additional articles were identified by manual search of references found in the primary articles. The data for clinical trials were collected by searching Clinical Trials.gov.Results: The liver has a tolerogenic nature in the intrahepatic milieu and its tumor microenvironment significantly affects tumor progression. The obstacles that reduce the efficacy of CAR-T therapy in solid tumors include a lack of specific tumor antigens, limited trafficking and penetration of CAR-T cells to tumor sites, and an immunosuppressive tumor microenvironment. To overcome these obstacles, several strategies have emerged. In addition, several strategies have been developed to manage the side effects of CAR-T, including enhancing the selectivity of CARs and controlling CAR-T activity. To date, no clinical trials of CAR-T therapy against HCC have been completed. However, preclinical studies in vitro and in vivo have shown potent antitumor efficacy. Glypican-3, mucin-1, epithelial cell adhesion molecule, carcinoembryonic antigen, and other targets are currently being studied.Conclusions: The application of CAR-T therapy for liver cancer is just beginning to be explored and more research is needed. However, we are optimistic that CAR-T therapy will offer a new approach for the treatment of liver cancers in the future.

Vascular endothelial growth factor attenuates hepatic sinusoidal capillarization in thioacetamide-induced cirrhotic rats

AIM: To investigate the effect of vascular endothelial growth factor (VEGF) transfection on hepatic sinusoidal capillarization. METHODS: Enhanced green fluorescent protein (EGFP)/ VEGF transfection was confirmed by immunofluorescence microscopy and immunohistoche-mistry both in primary hepatocytes and in normal liver. Cirrhotic rats were generated by thioacetamide (TAA) administration and then divided into a treatment group, which received injections of 400 μg of plasmid DNA encoding an EGFP- VEGF fusion protein, and a blank group, which received an equal amount of normal saline through the portal vein. The portal vein pressure was measured in the normal and cirrhotic state, in treated and blank groups. The average number of fenestrae per hepatic sinusoid was determined using transmission electron microscopy (TEM), while the relative abundance of VEGF transcripts was examined by Gene array. RESULTS: Green fluorescent protein was observed in the cytoplasms of liver cells under immunofluorescence microscopy 24 h after transfection with EGFP/VEGF plasmid in vitro. Staining with polyclonal antibodies against VEGF illustrated that hepatocytes expressedimmunodetectable VEGF both in vitro and in vitro. There were significant differences in the number of fenestrae and portal vein pressures between normal and cirrhotic rats (7.40 ± 1.71 vs 2.30 ± 1.16 and 9.32 ± 0.85 cmH2O vs 17.92 ± 0.90 cmH2O, P < 0.01), between cirrhotic and treated rats (2.30 ± 1.16 cmH2O vs 4.60 ± 1.65 and 17.92 ± 0.90 cmH2O vs 15.52 ± 0.93 cmH2O, P < 0.05) and between the treatment group and the blank group (4.60 ± 1.65 cmH2O vs 2.10 ± 1.10 cmH2O and 15.52 ± 0.93 cmH2O vs 17.26 ± 1.80 cmH2O, P < 0.05). Gene- array analysis revealed that the relative abundance of transcripts of VEGF family members decreased in the cirrhotic state and increased after transfection. CONCLUSION: Injection of a plasmid encoding VEGF through the portal vein is an effective method to induce the formation of fenestrae and decrease portal vein pressure in cirrhotic rats. Therefore, it may be a good choice for treating hepatic cirrhosis and portal hypertension.

COVID-19 or treatment associated immunosuppression may trigger hepatitis B virus reactivation: A case report

BACKGROUND Since the initial recognition of coronavirus disease 2019(COVID-19)in Wuhan,this infectious disease has spread to most areas of the world.The pathogenesis of COVID-19 is yet unclear.Hepatitis B virus(HBV)reactivation occurring in COVID-19 patients has not yet been reported.CASE SUMMARY A 45-year-old hepatitis B man with long-term use of adefovir dipivoxil and entecavir for antiviral therapy had HBV reactivation after being treated with methylprednisolone for COVID-19 for 6 d.CONCLUSION COVID-19 or treatment associated immunosuppression may trigger HBV reactivation.

Long noncoding RNA LINC01124 activates hepatocellular carcinoma cell proliferation, migration, and invasion by absorbing microRNA-1247-5p and overexpressing FOXO3

Long intergenic non-protein coding RNA 1124(LINC01124)has been identified as an important regulator of non-small-cell lung cancer.However,the expression and detailed role of LINC01124 in hepatocellular carcinoma(HCC)remain unestablished to date.Therefore,this study aimed to elucidate the role of LINC01124 in the aggressiveness of HCC cells and identify the underlying regulatory mechanism.Quantitative reverse transcriptase-polymerase chain reaction was performed to measure the expression of LINC01124 in HCC.Cell Counting Kit-8 assay,Transwell cell migration and invasion assays,and a xenograft tumor model were used to investigate the function of LINC01124 in HCC cells,and bioinformatics analysis,RNA immunoprecipitation,luciferase reporter assay,and rescue experiments were used to elucidate the underlying mechanisms.Herein,LINC01124 overexpression was confirmed in HCC tissues as well as cell lines.Further,the downregulation of LINC01124 decreased HCC cell proliferation,migration,and invasion in vitro,whereas the upregulation of LINC01124 triggered the opposite results.Additionally,LINC01124 ablation impaired tumor growth in vivo.Mechanistic analyses revealed that LINC01124 functions as a competing endogenous RNA to sponge microRNA-1247-5p(miR-1247-5p)in HCC cells.Moreover,forkhead box O3(FOXO3)was identified as a direct target of miR-1247-5p.FOXO3 was positively regulated by LINC01124 in HCC cells through the sequestration of miR-1247-5p.Finally,rescue assays revealed that the inhibition of miR-1247-5p or overexpression of FOXO3 reversed the effects of LINC01124 silencing on the HCC cell malignant phenotype.In summary,LINC01124 plays a tumor-promoting role in HCC by regulating the miR-1247-5p-FOXO3 axis.The LINC01124-miR-1247-5p-FOXO3 pathway may provide a foundation for the identification of alternative therapies for HCC.

PCDH17 increases the sensitivity of colorectal cancer to 5-fluorouracil treatment by inducing apoptosis and autophagic cell death

5-Fluorouracil(5-FU)is known as a first-line chemotherapeutic agent against colorectal cancer(CRC),but drug resistance occurs frequently and significantly limits its clinical success.Our previous study showed that the protocadherin 17(PCDH17)gene was frequently methylated and functioned as a tumor suppressor in CRC.However,the relationship between PCDH17 and 5-FU resistance in CRC remains unclear.Here,we revealed that PCDH17 was more highly expressed in 5-FU-sensitive CRC tissues than in 5-FU-resistant CRC tissues,and high expression of PCDH17 was correlated with high BECN1 expression.Moreover,this expression profile contributed to superior prognosis and increased survival in CRC patients.Restoring PCDH17 expression augmented the 5-FU sensitivity of CRC in vitro and in vivo by promoting apoptosis and autophagic cell death.Furthermore,autophagy played a dominant role in PCDH17-induced cell death,as an autophagy inhibitor blocked cell death to a greater extent than the pancaspase inhibitor Z-VAD-FMK.PCDH17 inhibition by siRNA decreased the autophagy response and 5-FU sensitivity.Mechanistically,we showed that c-Jun NH2-terminal kinase(JNK)activation was a key determinant in PCDH17-induced autophagy.The compound SP600125,an inhibitor of JNK,suppressed autophagy and 5-FU-induced cell death in PCDH17-reexpressing CRC cells.Taken together,our findings suggest for the first time that PCDH17 increases the sensitivity of CRC to 5-FU treatment by inducing apoptosis and JNK-dependent autophagic cell death.PCDH17 may be a potential prognostic marker for predicting 5-FU sensitivity in CRC patients.

The effects of genistein on transforming growth factor-β1-induced invasion and metastasis in human pancreatic cancer cell line Panc-1 in vitro

Background Pancreatic cancer is a devastating disease with the worst mortality rate. Therefore, a rational strategy for future drug development is critical. Genistein is a small, biologically active flavonoid that is found in high amounts in soy. This important compound supports a wide variety of biological activities, but is best known for its ability to inhibit cancer progression. Methods Transwell chamber assay was performed to determine the effect of genistein on the invasion of the human pancreatic cancer cell line Panc-1 induced by transforming growth factor-β1 （TGF-β1） in the different condition （5 ng/ml 24 hours and 10 ng/ml 48 hours）; Reverse transcription-polymerase chain reaction （RT-PCR） was used to estimate the mRNA levels of urinary plasminogen activator （uPA）, matrix metallopeptidase 2/9 （MMP-2/9）, Smad4, E-Cadherin and Vimentin; Western blotting was used to detect the protein levels of uPA, E-Cadherin, ERK1/2, P38 and P-P38, and the activity of MMP-2/9 protein were detected by gelatin zymography assay method. Cells structure was observed and analyzed by microscopy. Results Genistein can inhibit effectively TGF-β1-induced invasion and metastasis in Panc-1 by Transwell assay, which is through regulating the mRNA and protein expression of uPA and MMP2, but not MMP9 by RT-PCR / Western blotting, and is positively correlated with the concentration of genistein. At the same time, genistein also could improve the progress of epithelial-mesenchymal transition （EMT） via morphology observation using light microscopy / transmission electron microscopy （TEM）, which is mediated by the down-regulation of E-cadherin and the up-regulation of vimentin. Conclusions TGF-β1 mediates EMT process via numerous intracellular signal transduction pathways. The potential molecular mechanisms are all or partly through Smad4-dependent and -independent pathways （p38 MAPK） to regulate the antitumor effect of genistein.

Restrictive versus liberal fluid therapy for major abdominal surgery: is the evidence strong?

We have read with great interest the original study by Myles et al. (1), in which the authors claimed a restrictive fluid regimen during and up to 24 hours after major abdominal surgery was associated with higher rate of acute kidney injury. This international, randomized, assessor-blinded trail compared the following outcomes between restrictive and liberal intravenous-fluid regimen: disability-free survival at 1 year, acute kidney injury at 30 days, renal-replacement therapy at 90 days, septic complications, surgical-site infection, and death. The perioperative fluid management is of great clinical significance, the results demonstrated by Myles et al. provide high-level evidence with dedicate design and adequate case volume, serving as potential reference to guidelines of surgical patient care. Shortly after its online publication, the article has drawn tremendous attention and caused extensive discussion among surgeons world-wide.

Comparative long-term outcomes for pancreatic volume change, nutritional status, and incidence of new-onset diabetes between pancreatogastrostomy and pancreatojejunostomy after pancreaticoduodenectomy

Background:The difference in volume change in a pancreatic remnant according to the type of pancreaticoenterostomy after pancreaticoduodenectomy(PD)for long-term follow-up is unknown.Also,there are few studies that evaluate the difference in general nutritional status and pancreatic endocrine function,including new-onset diabetes mellitus(NODM)depending on the type of pancreaticoenterostomy.This study aimed to compare serial pancreatic volume changes in pancreatic remnants between pancreatogastrostomy(PG)and pancreatojejunostomy(PJ)after PD and to evaluate the difference in general nutritional status and incidence of NODM between PG and PJ.Methods:This study enrolled 115 patients who had survived for more than 3 years after PD.They were divided into the PG group and the PJ group.Their clinicopathologic factors were collected and analyzed.We calculated serial pancreas volume and pancreatic duct size precisely from preoperative stage to 5 years after surgery by image-processing software specifically designed for navigation and visualization of multimodality and multidimensional images.Consecutive changes of albumin and body mass index(BMI)as related to general nutritional status were compared between the PG and PJ groups.To evaluate the incidence and risk factors of NODM following PD,subgroup analysis was performed in 88 patients who did not have diabetes preoperatively.Results:Most patient demographics were not significantly different between the PG group(n=45)and PJ group(n=70).There was no significant difference in volume reduction between the groups from postoperative 1 month to 5 years(PG group?18.21±14.66 mL versus PJ group?14.43±13.05 mL,P=0.209).But there was a significant difference in increased pancreatic duct size between the groups from postoperative 1 month to 5 years(PG group 1.66±2.20 mm versus PJ group 0.54±1.54 mm,P=0.007).There was no significant difference in the increase of total serum albumin between the groups for 5 years after surgery(PG group 0.51±0.47 g/dL,14.3%versus PJ group 0.42±0.60 g/dL,11.3%,P=0.437).There was also no significant difference in BMI decrease between the groups(PG group?1.13±3.12,?4.9%versus PJ group?1.97±2.01,?8.7%,P=0.206).On the whole,NODM was diagnosed in 19 patients out of the 88 patients(21.6%)who did not have DM preoperatively.The incidence of NODM was not significantly different between the groups(PG group 21.6%versus PJ group 21.5%,P=0.995).In addition,pancreaticoenterostomy was not an independent risk factor for NODM by logistic regression analysis(odds ratio,0.997,95%CI:0.356–0.2.788,P=0.995).No other risk factors for NODM were found.Conclusions:PG and PJ following PD induced similar pancreatic volume reduction during long-term follow-up.There was no difference in general nutritional status or incidence of NODM between the groups after PD.

A versatile ligand-assisted cooperative template method to synthesize multi-shelled mesoporous hollow metal hydroxide and oxide nanospheres as catalytic reactors

Nowadays,multi-shelled mesoporous hollow metal oxide nanospheres have drawn a lot of attention due to their large internal space,nanometer scaled shell thickness,high specific surface area and well-defined mesopores,of which unique nanostructure endows metallic oxides with enhanced properties.In this thesis,we have studied and proposed a versatile ligand-assisted cooperative template method to synthesize multi-shelled mesoporous hollow metal hydroxides and oxides nanospheres,in which silica nanospheres act as sacrificial templates and the coordination interaction between metal ions and surfactant can be cooperatively amplified by using chelating ligand(ascorbic acid)as a co-template.The synthesized metal hydroxides and oxides nanospheres possess stable hollow structure,uniform spherical morphology and tunable diameter from 270 to 690 nm.All the multi-shelled mesoporous hollow metal hydroxide and metal oxide nanospheres exhibit high surface areas(up to 640 m^(2)/g).The obtained Au nanoparticles loaded composited nanospheres exhibit excellent reactivity for solvent-free aerobic oxidation of ethylbenzene with high activity(28.2%)and selectivity(87%).