First report on establishment and characterization of the extrahepatic cholangiocarcinoma sarcoma cell line CBC2T-2.

BACKGROUND: Extrahepatic cholangiocarcinoma sarcoma is extremely rare in clinical practice. These cells consist of both epithelial and mesenchymal cells. Patient-derived cell lines that maintain tumor characteristics are valuable tools for studying the molecular mechanisms associated with carcinosarcoma. However, cholangiocarcinoma sarcoma cell lines are not available in cell banks. AIM: To establish and characterize a new extrahepatic cholangiocarcinoma sarcoma cell line, namely CBC2T-2. METHODS: We conducted a short tandem repeat (STR) test to confirm the identity of the CBC2T-2 cell line. Furthermore, we assessed the migratory and invasive properties of the cells and performed clonogenicity assay to evaluate the ability of individual cells to form colonies. The tumorigenic potential of CBC2T-2 cells was tested in vivo using non-obese diabetic/severe combined immunodeficient (NOD/SCID) mice. The cells were injected subcutaneously and tumor formation was observed. In addition, immunohistochemical analysis was carried out to examine the expression of epithelial marker CK19 and mesenchymal marker vimentin in both CBC2T-2 cells and xenografts. The CBC2T-2 cell line was used to screen the potential therapeutic effects of various clinical agents in patients with cholangiocarcinoma sarcoma. Lastly, whole-exome sequencing was performed to identify genetic alterations and screen for somatic mutations in the CBC2T-2 cell line. RESULTS: The STR test showed that there was no cross-contamination and the results were identical to those of the original tissue. The cells showed round or oval-shaped epithelioid cells and mesenchymal cells with spindle-shaped or elongated morphology. The cells exhibited a high proliferation ratio with a doubling time of 47.11 h. This cell line has migratory, invasive, and clonogenic abilities. The chromosomes in the CBC2T-2 cells were polyploidy, with numbers ranging from 69 to 79. The subcutaneous tumorigenic assay confirmed the in vivo tumorigenic ability of CBC2T-2 cells in NOD/SCID mice. CBC2T-2 cells and xenografts were positive for both the epithelial marker, CK19, and the mesenchymal marker, vimentin. These results suggest that CBC2T-2 cells may have both epithelial and mesenchymal characteristics. The cells were also used to screen clinical agents in patients with cholangiocarcinoma sarcoma, and a combination of paclitaxel and gemcitabine was found to be the most effective treatment option. CONCLUSION: We established the first human cholangiocarcinoma sarcoma cell line, CBC2T-2, with stable biogenetic traits. This cell line, as a research model, has a high clinical value and would facilitate the understanding of the pathogenesis of cholangiocarcinoma sarcoma.

[In-situ Remediation Effect of Cadmium-polluted Agriculture Land Using Different Amendments Under Rice-wheat Rotation].

Exploring the effects of one-time amendment treatments on cadmium (Cd)-contaminated farmland soils is beneficial for providing a theoretical basis to effectively prevent Cd pollution in farmland soils and ensure the safe production of crops. Five amendments, including straw biochar, fly ash, sepiolite, white marble powder, and shale (particle size <0.2 mm, application rate 2.25 kg·m-2), were applied to the Cd-contaminated farmland soils. The soil nutrients, pH, soil available Cd, and Cd chemical forms in the soils and grain Cd concentration in the planted crops were determined to investigate the effects and persistence of one-time applications of the five amendments. The results showed that:① the application of the five amendments had little effect on soil nutrient content, but all of them could increase soil pH. Amendment treatments improved the transfer of Cd from the acid extraction fraction to residue fraction and further reduced the Cd availability in the soil. The decreasing amplitudes of straw biochar and white marble powder soil conditioner were 20.42%-22.53%, which was higher than those in the other treatments. ② The grain Cd concentrations in rice and wheat were significantly decreased under the amendment treatments with the decreasing amplitudes of 19.88%-48.77% and 5.06%-24.00%, respectively. The Cd concentrations in rice grains under the treatments of straw biochar, fly ash, and white marble powder soil conditioner were 0.195, 0.196, and 0.223 mg·kg-1, respectively, which were lower than those under the other treatments and were close to or approached the National Standard of Food Safety(GB 2762-2017)(0.2 mg·kg-1). ③ The immobilization effects on Cd in farmland soils were decreasing with time under one-time application of the amendments. The available Cd concentrations in the soil and Cd concentrations in crop grains were still lower than those in the control after three rounds of rice-wheat rotation. The straw biochar and white marble powder soil conditioner had a good and long-term effect on reducing Cd availability in soils and Cd concentrations in crop grain, making them ideal materials for safe production in Cd-contaminated soils.

The inflammatory cytokine profiles and ocular biometric characteristics of primary angle-closure glaucoma.

OBJECTIVE: To investigate the pathogenesis of primary angle-closure disease (PACG) by measuring the anatomical structures of the anterior and posterior segments of the eye and inflammatory markers in the peripheral blood. METHODS: This case-control study enrolled patients diagnosed with acute PACG (APACG) and chronic PACG (CPACG). It also enrolled control subjects without PACG. The anterior and posterior anatomical features were measured in all study participants. The levels of interleukin (IL)-6, tumour necrosis factor-α and the neutrophil-to-lymphocyte ratio (NLR) in the peripheral blood were measured. RESULTS: This study analysed a total of 99 eyes: 34 eyes from 34 patients with APACG, 28 eyes from 28 patients with CPACG and 37 eyes from 37 control patients with senile cataract. The axis length, corneal diameter, anterior chamber depth and anterior chamber volume were significantly smaller in the APACG and CPACG groups compared with the controls. The level of IL-6 in the peripheral blood of patients with PACG was significantly lower than that of the controls. The NLR in the peripheral blood of patients with PACG was significantly greater than that of the controls. CONCLUSIONS: Changes in the ocular anatomy and some inflammatory markers might be involved in the pathogenesis of PACG.

[Effects of Different Amendments on Cadmium Accumulation in Rice Safety in Cadmium-Contaminated Farmland Under Two Flooding Treatments].

Cadmium (Cd) accumulation in rice has become a serious public concern; thus, it is important to find an effective approach to reducing Cd accumulation in rice grains to ensure food safety. To investigate the effects of different amendments on Cd accumulation in rice in Cd-contaminated farmland under different flooding treatments, a field experiment with three amendments (jade powder, biochar, and fly ash) and two flooding treatments (intermittent flooding and flooding throughout the whole growth period) was conducted. The results showed that:① without amendment application, the soil pH significantly increased, whereas the soil available Cd concentration decreased by 3.81%-17.27% and 2.25%-6.74% with the treatments of flooding throughout the whole growth period and intermittent flooding, respectively. Additionally, the immobilizing efficiency of the treatment of flooding throughout the whole growth period was better than that of intermittent flooding; ② under different flooding treatments, amendment application improved soil pH, resulting in a decrease in the soil available Cd concentration along with an increase in the residual Cd concentration. Under the treatment of intermittent flooding, the soil pH increased by 0.19-2.20 units, and the soil available Cd concentration and Cd concentration in rice grains decreased by 4.72%-22.68% and 2.60%-75.75%, respectively, with the application of different amendments. Under the treatment of flooding throughout the whole growth period, the application of different amendments decreased the soil available Cd concentration and Cd concentration in rice grains by 5.06%-36.63% and 13.28%-77.01%, respectively. The immobilizing efficiency in both flooding treatments was jade powder > biochar > fly ash. ③ Under different flooding treatments, the application of amendments significantly reduced the soil available Cd concentration and Cd concentration in rice grains. Among the three amendments, jade powder showed the best capacity of immobilizing efficiency with the treatment of flooding throughout the whole growth period; the soil Cd reduction rates were 36.63% and 25.16%, and the Cd concentrations in rice grains were 0.058 and 0.170 mg·kg-1 in 2019 and 2020, respectively. The Cd concentrations in rice grains were within the limitation of the National Food Hygienic Standard of China. Therefore, combining flooding throughout the whole growth period with jade powder can be considered as an ideal strategy for ensuring rice safety in Cd-contaminated farmland.

A case report of pyopneumopericardium following bungee jumping in a patient with tuberculosis.

RATIONALE: Pyopneumopericardium related to bungee jumping is a rare occurrence in the current antibiotic era. We present a case of esophagus-seeded Streptococcus sanguinis pyopneumopericardium in a young man with tuberculosis who had just completed bungee jumping. PATIENT CONCERN: A 27-year-old man was hospitalized with a 1-day history of fever, chest tightness, and intermittent sharp chest pain after bungee jumping for the first time. DIAGNOSES: Clinical examinations, thoracentesis, and pericardiocentesis revealed pyopneumopericardium, pyopneumomediastinum, and suppurative pleurisy secondary to bungee-jumping-related traumas. Pericardial fluid cultures were positive for S sanguinis, and Mycobacterium tuberculosis complex genetic test was positive in both sputum and pleural effusion. INTERVENTIONS: The patient improved with drainage and comprehensive antimicrobial therapy. OUTCOMES: The patient developed constrictive pericarditis and underwent pericardiectomy after 6 months of anti-tuberculosis treatment. During the 6-month follow-up after surgery, he recovered uneventfully. LESSONS: This case adds to the long list of bungee-jumping complications. Early diagnosis to initiate appropriate therapy is critical for pyopneumopericardium patients to achieve good outcomes.

Oral Microbiota Community Dynamics Associated With Oral Squamous Cell Carcinoma Staging.

Oral squamous cell carcinoma (OSCC) is a highly aggressive cancer and the fourth leading malignancy among males in Taiwan. Some pathogenic bacteria are associated with periodontitis and oral cancer. However, the comprehensive profile of the oral microbiome during the cancer's progression from the early stage to the late stage is still unclear. We profiled the oral microbiota and identified bacteria biomarkers associated with OSCC. The microbiota of an oral rinse from 51 healthy individuals and 197 OSCC patients at different stages were investigated using 16S rRNA V3V4 amplicon sequencing, followed by bioinformatics and statistical analyses. The oral microbiota communities from stage 4 patients showed significantly higher complexity than those from healthy controls. The populations also dynamically changed with the cancer's progression from stage 1 to stage 4. The predominant phyla in the oral samples showed variation in the relative abundance of Fusobacteria, Bacteroidetes, and Actinobacteria. The abundance of Fusobacteria increased significantly with the progression of oral cancer from the healthy controls (2.98%) to OSCC stage 1 (4.35%) through stage 4 (7.92%). At the genus level, the abundance of Fusobacterium increased, while the number of Streptococcus, Haemophilus, Porphyromonas, and Actinomyces decreased with cancer progression. Fusobacterium periodonticum, Parvimonas micra, Streptococcus constellatus, Haemophilus influenza, and Filifactor alocis were associated with OSCC, and they progressively increased in abundance from stage 1 to stage 4. The abundances of Streptococcus mitis, Haemophilus parainfluenzae, and Porphyromonas pasteri were inversely associated with OSCC progression. We selected a bacterial marker panel of three bacteria (upregulated F. periodonticum, down-regulated S. mitis, and P. pasteri), which had an AUC of 0.956 (95% CI = 0.925-0.986) in discriminating OSCC stage 4 from the healthy controls. Furthermore, the functional prediction of oral bacterial communities showed that genes involved in carbohydrate-related metabolism, such as methane metabolism, and energy-metabolism-related parameters, such as oxidative phosphorylation and carbon fixation in photosynthetic organisms, were enriched in late-stage OSCC, while those responsible for amino acid metabolism, such as folate biosynthesis and valine, leucine, and isoleucine biosynthesis, were significantly associated with the healthy controls. In conclusion, our results provided evidence of oral bacteria community changes during oral cancer progression and suggested the possibility of using bacteria as OSCC diagnostic markers.

Synthesis and characterization of Tamoxifen citrate modified reduced graphene oxide nano sheets for breast cancer therapy.

Theranostic agents are of immense consideration in the current generation nanomedicine. In this study, we have developed a facile approach for the fabrication of Tamoxifen citrate modified nanosized reduced graphene oxide (nano-rGO) with more stability and low cytotoxicity. The prepared nano-rGO sheets were characterized using HR-TEM and AFM imaging techniques. Further, the cytotoxicity was assessed using MTT assay on female BALB/c nude mice MCF-7 cell lines. In addition, by means of continuous-wave near-infrared laser, cancer cells in vivo were significantly ablated because of the photothermal effect stimulated by tamoxifen modified nano-rGO. These results indicated that the prepared tamoxifen modified nano-rGO has the ability to apply in the photothermal therapy of breast cancers. Consequently, further exploration of photothermal therapeutics is desirable for the synthesis of novel nano materials with additional functionalities.

Stevioside induced cytotoxicity in colon cancer cells via reactive oxygen species and mitogen-activated protein kinase signaling pathways-mediated apoptosis.

The role of mitogen-activated protein kinase (MAPK) signaling pathways in cell growth and differentiation has been well established. The present study aimed to investigate the anti-proliferative effect of stevioside on human colon cancer HT-29 cells. Additionally, the effect of stevioside on cell cycle arrest and MAPK signaling pathways in HT-29 cells was explored. Stevioside was observed to significantly inhibit cancer cell growth at a dose of 5 µM at 48 and 72 h. A dose-dependent increase in the apoptosis rate was observed with cell cycle arrest at G2/M phase. In addition, caspase-9 and caspase-3 activity also increased. An increase in reactive oxygen species (ROS) production and a decrease in the mitochondrial membrane potential indicated that the mitochondrial-mediated intrinsic pathway is responsible for apoptotic activity. These results were additionally verified by the elevated expression level of phosphorylated p38 and extracellular signal-regulated kinase mitogen-activated protein kinases (MAPKs). Additionally, by inhibiting ROS production and MAPK activation, the antiproliferative effect of stevioside was suppressed, confirming the hypothesis that ROS and MAPK proteins induce apoptosis in human colon cancer HT-29 cells.

The rapid antidepressant effect of ketamine in rats is associated with down-regulation of pro-inflammatory cytokines in the hippocampus.

OBJECTIVES: Active inflammatory responses play an important role in the pathogenesis of depression. We hypothesized that the rapid antidepressant effect of ketamine is associated with the down-regulation of pro-inflammatory mediators. METHODS: Forty-eight rats were equally randomized into six groups (a control and five chronic unpredictable mild stress (CUMS) groups) and given either saline or 10 mg/kg ketamine, respectively. The forced swimming test was performed, and the hippocampus was subsequently harvested for the determination of levels of interleukin (IL)-1ß, IL-6, tumour necrosis factor-α (TNF-α), indoleamine 2,3-dioxygenase (IDO), kynurenine (KYN), and tryptophan (TRP). RESULTS: CUMS induced depression-like behaviours and up-regulated the hippocampal levels of IL-1ß, IL-6, TNF-α, IDO, and the KYN/TRP ratio, which were attenuated by a sub-anaesthetic dose of ketamine. CONCLUSION: CUMS-induced depression-like behaviours are associated with a reduction in hippocampal inflammatory mediators, whereas ketamine's antidepressant effect is associated with a down-regulation of pro-inflammatory cytokines in the rat hippocampus.

Nontuberculous mycobacterial osteomyelitis.

Osteomyelitis caused by nontuberculous mycobacteria (NTM) can have severe consequences and a poor prognosis. Physicians therefore need to be alert to this condition, especially in immunocompromised patients. Although the pathogenesis of NTM osteomyelitis is still unclear, studies in immunodeficient individuals have revealed close relationships between NTM osteomyelitis and defects associated with the interleukin-12-interferon-γ-tumor necrosis factor-α axis, as well as human immunodeficiency virus infection, various immunosuppressive conditions, and diabetes mellitus. Culture and species identification from tissue biopsies or surgical debridement tissue play crucial roles in diagnosing NTM osteomyelitis. Suitable imaging examinations are also important. Adequate surgical debridement and the choice of appropriate, combined antibiotics for long-term anti-mycobacterial chemotherapy, based on in vitro drug susceptibility tests, are the main therapies for these bone infections. Bacillus Calmette-Guerin vaccination might have limited prophylactic value. The use of multiple drugs and long duration of treatment mean that the therapeutic process needs to be monitored closely to detect potential side effects. Adequate duration of anti-mycobacterial chemotherapy together with regular monitoring with blood and imaging tests are key factors determining the recovery outcome in patients with NTM osteomyelitis.

High expression of 23 kDa protein of augmenter of liver regeneration (ALR) in human hepatocellular carcinoma.

BACKGROUND: Augmenter of liver regeneration (ALR) is an important polypeptide that participates in the process of liver regeneration. Two forms of ALR proteins are expressed in hepatocytes. Previous data have shown that ALR is essential for cell survival and has potential antimetastatic properties in hepatocellular carcinoma (HCC). AIMS: The study aimed to evaluate the expression levels of two forms of ALR proteins in HCC and their possible significance in HCC development. METHODS: Balb/c mouse monoclonal antibody against ALR protein was prepared in order to detect the ALR protein in HCC by Western blotting and immunohistochemistry. ALR mRNA expression levels were measured by real-time polymerase chain reaction in HCC tissues and compared to paracancerous liver tissues in 22 HCC patients. RESULTS: ALR mRNA expression in HCC liver tissues (1.51×10(6) copies/µL) was higher than in paracancerous tissues (1.04×10(4) copies/µL). ALR protein expression was also enhanced in HCC liver tissues. The enhanced ALR protein was shown to be 23 kDa by Western blotting. Immunohistochemical analysis showed that the 23 kDa ALR protein mainly existed in the hepatocyte cytosol. CONCLUSION: The 23 kDa ALR protein was highly expressed in HCC and may play an important role in hepatocarcinogenesis.

Liver tuberculosis presenting as an uncommon cause of pyrexia of unknown origin: positron emission tomography/computed tomography identifies the correct site for biopsy.

OBJECTIVE: To identify the correct site to biopsy in a case of pyrexia of unknown origin (PUO) caused by hepatic tuberculosis (TB). CLINICAL PRESENTATION AND INTERVENTION: A 58-year-old man who developed hepatic TB presented with PUO. Ultrasonography (US) and computed tomography (CT) of the abdomen showed only calcifications in the liver, and positron emission tomography (PET)/CT showed diffuse increased metabolic activity in addition to focal areas of increased activity. A diagnosis of hepatic TB was confirmed by histological examination of liver tissues and interferon-γ release assays (IGRAs of T-SPOT/TB). The patient was treated with 4 anti-tubercular therapies (rifampicin, isoniazid, ethambutol and pyrazinamide). At the 3-month follow-up, the patient was disease free as confirmed by abdominal US. CONCLUSION: PET/CT was helpful in identifying a site to biopsy that led to the correct diagnosis.

Diagnostic value of fluorine-18 fluorodeoxyglucose positron emission tomography/computed tomography in patients with fever of unknown origin.

BACKGROUND: While fever of unknown origin (FUO) remains a challenging problem in clinical practice, fluorine-18 fluorodeoxyglucose ((18)F-FDG) positron emission tomography/computed tomography (PET/CT) has been considered helpful in diagnosing its cause. The present study is set to evaluate the diagnostic value of PET/CT for patients with FUO. METHODS: We analyzed the records of 48 patients with FUO (34 men and 14 women; mean age of 57-year-old with a range between 24- and 82-year-old). The patients were examined by (18)F-FDG PET/CT and the results were compared to a final diagnosis that was established by additional procedures. RESULTS: A final diagnosis was established for 36 patients (75%). Among them, 15 patients had infectious diseases, 12 patients had malignancies, and 9 patients had non-infectious inflammatory diseases. Thirty-two abnormal PET/CT results correctly revealed the source of fever (true-positives). Abnormal PET/CT results were considered false-positives for 8 patients without diagnoses. Normal PET/CT results in 4 patients with no diagnoses were classified as true-negatives. Four patients with normal PET/CT results with diagnosed cause for FUO were considered false-negatives. Therefore, PET/CT had a positive predictive value of 80%, a negative predictive value of 50%, a sensitivity of 89%, and a specificity of 33% in patients with FUO. CONCLUSIONS: Our study demonstrated that FDG-PET/CT is a valuable imaging tool for the identification of the etiology in patients with FUO. The results suggest that this procedure may be considered as a second-line test, especially when conventional structural imaging was normal or unable to distinguish lesions from benign and malignant.

Expression level of augmenter of liver regeneration in patients with hepatic failure and hepatocellular carcinoma.

BACKGROUND: Augmenter of liver regeneration (ALR) is an important polypeptide in the process of liver regeneration. This study aimed to determine the expression level of ALR in different liver diseases and its significance. METHODS: We prepared murine polyclonal antibody against ALR protein from Balb/C mice and purified the IgG fraction, which specifically combined to ALR protein as shown by Western blotting. Serum ALR levels in patients with hepatocellular carcinoma (HCC), hepatic failure (HF), chronic hepatitis B, and healthy persons were compared by ELISA. ALR mRNA expression levels in liver tissues in some of these patients were also compared by real-time RT-PCR. Immunohistochemical analysis was carried out on HF and HCC liver tissues. RESULTS: Different serum ALR levels foreshowed completely different prognoses in 18 HF patients. Higher ALR levels were noted in 6 improved patients (1613.5+/-369.6 pmol/ml) than in 12 deteriorating patients (462.3+/-235.8 pmol/ml). Similar levels were found in 20 HCC patients (917.9+/-332.7 pmol/ml), 24 chronic hepatitis B patients (969.2+/-332.5 pmol/ml) and 10 healthy persons (806.9+/-240.8 pmol/ml). ALR mRNA levels in HCC liver tissues [10E6.24 (1.74X10(6)) copies/µl] were much higher than in those of HF patients receiving orthotopic liver transplantation [10E3.45 (2.82X10(3)) copies/µl] or in healthy liver tissues [10E4.31 (2.04X10(4)) copies/µl]. In immunohistochemical analysis, positive immunostaining in HCC liver tissue was more intense than that in HF liver tissue. CONCLUSION: Serum ALR level is helpful in estimating the survival time of patients with HF, and ALR may play an important role in hepatocarcinogenesis.

Effects of blood-activating and stasis-removing drugs combined with VEGF gene transfer on angiogenesis in ischemic necrosis of the femoral head.

OBJECTIVE: To observe the promoting effects of blood-activating and stasis-removing Chinese drugs combined with vascular endothelial growth factor (VEGF) gene transfer on angiogenesis in ischemic necrosis of the femoral head. METHODS: Forty Japanese giant-ear rabbits were randomly divided into a control group, a model group, a Chinese drug group, a gene group, and a combined group. After 8 weeks of treatment, the rate of VEGF positive cell expression in the synovium of the femoral head was measured using the immunohistochemical method, and the number of blood vessels in the femoral head was measured by digital subtraction angiography. RESULTS: The rate of VEGF positive cell expression in the model group was significantly lower than that in the Chinese drug group (P < 0.05) and very significantly lower than those in other groups (P < 0.01); but in the combined group it was significantly higher than in the Chinese drug group (P < 0.05). The differences in the number of blood vessels in area A between the model group and other groups were not statistically significant. However, in area B, the number of blood vessels significantly increased in the control group, the gene group and the combined group as compared with the model group (P < 0.05), and in the combined group the number of blood vessels was significantly more than in the gene group (P < 0.05); but in the Chinese drug group it was not significantly different than the model group (P > 0.05). CONCLUSION: Either the blood-activating and stasis-removing Chinese drugs or VEGF gene transfer can promote the angiogenesis and building of collateral circulation for femoral head ischemic necrosis, and the combined therapy with Chinese drugs or VEGF gene transfer may show a better therapeutic effect. The present study provides an experimental basis for clinical application of the combined therapy with the blood-activating and stasis-removing Chinese drugs and VEGF gene transfer.

Establishment of highly differentiated immortalized human hepatocyte line with simian virus 40 large tumor antigen for liver based cell therapy.

Acute liver failure and metabolic liver disorder animal models have demonstrated that hepatocytes transplanted into the liver or spleen survive and participate in the liver repopulation process, and recent studies have documented the usefulness of hepatocyte transplantation in humans. However, despite the promising cell therapy, there are still many restrictions, such as the shortage of donor human livers and the limited lifespan and the functional insufficiency of primary cultured hepatocytes. The immortalized and highly differentiated human hepatocyte could provide an unlimited supply of transplantable cells. In this study, we established an efficient and highly differentiated immortalized human hepatocyte line for bioartificial liver and hepatocyte transplantation research. Hepatocytes isolated from the liver of a 25 year old, brain dead male were transfected with pcDNA3.1 (-) recombinant plasmid containing the genes encoding simian virus 40 (SV40) large tumor antigen. One of the hepatocyte clones, HepLL, displayed highly differentiated liver functions with immortalized characteristics and was selected with a 700-300 microg/ml of G418 technique in 42 days. To characterize this immortalized cell line for cell therapy in the near future, HepLL cells were studied with immunohistochemistry, reverse transcription-polymerase chain reactions, immunoblotting, and tumorigenicity tests. The results revealed that HepLL cells displayed morphologic characteristics of liver parenchymal cells, secreted albumin, synthesized urea and glycogen, and expressed liver enriched functional markers, but there were no tumorigenic qualities after transplantation into severe combined immunodeficiency mice. Thus this immortalized human hepatocyte line is expected to be a useful tool for studying the functions of differentiated human hepatocyte and a promising strategy to resolve the shortages of donor organs and the limits of primary human hepatocyte for transplantation and bioartificial liver support systems.