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The correlation between chronic hepatitis B virus (HBV) infection and hepatocellular carcinoma (HCC) has been well-established. But the roles of viral factor remain uncertain. Only HBV X gene and nonsense mutations of S gene (C-terminal truncation of HBV surface protein) have been demonstrated to have transforming activity. Whether they play a significant role in hepatocarcinogenesis is still uncertain. Twenty-five HBV-related HCC patients were positive for hepatitis B core antigen (HBcAg) in the cancerous parts of their HCC liver tissues by immunohistochemistry studies, and had available tissue for whole HBV genome sequence analysis. The results were compared with 25 gender and age-matched HBcAg negative HCCs. Plasmids encoding HBV S gene nonsense mutations identified from HBcAg (+) HCC tissue were constructed to investigate their cell proliferation, transformation activity and the oncogenic potentials by xenograft study and in vivo migration assay. HBcAg (+) HCC patients were significantly associated with cirrhosis and small tumor size (≦2 cm) when compared with HBcAg (-) HCC patients. Southern blot analyses revealed freely replicative forms of HBV in the cancerous parts of HBcAg(+) HCC. Three nonsense mutations of S gene (sL95*, sW182*, and sL216*) were identified in the HBcAg(+) HCC tumor tissues. sW182* and sL216* were recurrently found in the 25 HBcAg (-) HCC tumor tissue, too. Functional studies of the above 3 non-sense mutations all demonstrated higher cell proliferation activities and transformation abilities than wild type S, especially sW182*. Tumorigenicity analysis by xenograft experiments and in vitro migration assay showed potent oncogenic activity of sW182* mutant. This study has demonstrated potent oncogenic activity of nonsense mutations of HBV S gene, suggesting they may play an important role in hepatocarcinogenesis.

A single-tube multiprobe real-time PCR assay for simultaneous detection of Entamoeba histolytica and Entamoeba dispar was developed. One primer pair with 2 species-specific probes was designed based on new SSU RNA regions of the ribosomal DNA-containing episome. The sensitivity is 1 parasite per milliliter of feces and thus superior to the conventional nested PCR and comparable to other published real-time PCR protocols. The applicability for clinical diagnosis was validated with 218 stool specimens from patients. A total of 51 E. histolytica and 39 E. dispar positive samples was detected by the multiprobe real-time PCR compared to 39 and 22 by routine nested PCR diagnosis. The detection rate of Entamoeba species for the multiprobe real-time PCR assays was significantly higher than the nested PCR (40.8% vs. 28.0%, P < 0.01). The test did not show cross reactivity with DNA from Entamoeba moshkovskii, Giardia lamblia, Cryptosporidium sp., Escherichia coli, or other nonpathogenic enteric parasites. The multiprobe real-time PCR assay is simple and rapid and has high specificity and sensitivity. The assay could streamline the laboratory diagnosis procedure and facilitate epidemiological investigation.

To explore the feasibility of computer-aided monitoring of the workflow in surgical pathology. We collected 5-year data about computer-aided monitoring of the workflow in surgical pathology and analyzed the four subprocesses in the surgical pathologic process: 1) from arranging surgical pathology examination to receipt of the examination sheet and sample by the laboratory; 2) from receipt of the sample to issuance of the pathology report; 3) from issuance of the pathology report to automatic computer forwarding of positive pathology reports by e-mail to the physician who ordered the examination; 4) from receipt of the positive report by the physician to his/her response of acknowledging receipt. A total 115,648 surgical pathological cases were reviewed in this study. The overdue rate of delivery of samples was 0.82 %. The most common cause (62.92 %) of overdue delivery was clinicians in the outpatient department arranging for the examination more than 1 day in advance of specimen collection. The cumulative rates of report completion within 1, 2, 3, 4 and 5 work days were 12.82 %, 53.56 %, 86.42 %, 95.90 % and 98.85 %, respectively. The rate of overdue reporting was 1.15 % over the 5-year study. The most common cause (56.30 %) of overdue reporting was case complexity. The learning time for adapting this subprocess of report issuance was 7 months. There were 12,151 positive reports (10.51 % of all cases) that required automatic computer forwarding to the physicians’ e-mail boxes. A total of 113 cases (0.93 %) failed in automatic computer forwarding during the 5-year period. The learning time for constructing a stable automatic computer forwarding system was 2.5 years. Of the 12,038 reports successfully forwarded, 10,107 (83.96 %) were received by physicians and acknowledged by automated receipt within 120 h, and the other 1,931 (16.04 %) showed no response within 120 h. The major reason for an overdue reply was that the physicians did not check their e-mail boxes (94.89 %). We used a preliminary computer-aided system to monitor the workflow in surgical pathology. This system might be used as one of the methods of quality assurance in surgical pathology.

Background & Aims The correlation between chronic hepatitis B virus (HBV) infection and hepatocellular carcinoma (HCC) has been well-established. But the roles of viral factor remain uncertain. Only HBV X gene and nonsense mutations of S gene (C-terminal truncation of HBV surface protein) have been demonstrated to have transforming activity. Whether they play a significant role in hepatocarcinogenesis is still uncertain. Methods Twenty-five HBV-related HCC patients were positive for hepatitis B core antigen (HBcAg) in the cancerous parts of their HCC liver tissues by immunohistochemistry studies, and had available tissue for whole HBV genome sequence analysis. The results were compared with 25 gender and age-matched HBcAg negative HCCs. Plasmids encoding HBV S gene nonsense mutations identified from HBcAg (+) HCC tissue were constructed to investigate their cell proliferation, transformation activity and the oncogenic potentials by xenograft study and in vivo migration assay. Results HBcAg (+) HCC patients were significantly associated with cirrhosis and small tumor size ([lE]2 cm) when compared with HBcAg (-) HCC patients. Southern blot analyses revealed freely replicative forms of HBV in the cancerous parts of HBcAg(+) HCC. Three nonsense mutations of S gene (sL95*, sW182*, and sL216*) were identified in the HBcAg(+) HCC tumor tissues. sW182* and sL216* were recurrently found in the 25 HBcAg (-) HCC tumor tissue, too. Functional studies of the above 3 non-sense mutations all demonstrated higher cell proliferation activities and transformation abilities than wild type S, especially sW182*. Tumorigenicity analysis by xenograft experiments and in vitro migration assay showed potent oncogenic activity of sW182* mutant. Conclusions This study has demonstrated potent oncogenic activity of nonsense mutations of HBV S gene, suggesting they may play an important role in hepatocarcinogenesis.

Aim： To investigate the effect of isochaihulactone （also known as K8）, a lignan compound of Bupleurum scorzonerifolium, on H2O2- induced cytotoxicity in neuronally differentiated PC12 cells （nPC12）. Methods： Viability of neuronal PC12 cells was measured using MTT assay. Protein expression was determined by Western blot. Apoptotic cells was determined using TUNEL assay. D-galactose aging mice were used as a model system to study the anti-oxidant effects of isochaihulactone in vivo. Results： Pretreatment with isochaihulactone （5-10 μmol/L） increased cell viability and decreased membrane damage, generation of reactive oxygen species and degradation of poly （ADP-ribose） polymerase in H2O2-treated nPC12 cells and also decreased the expression of cyclooxygenase-2, via downregulation of NF-kappaB, resulting in a decrease in lipid peroxidation. The results suggest that isochaihulactone is a potential antioxidant agent. In a murine aging model, in which chronic systemic exposure to D-galactose （D-gal） causes the acceleration of senescence, administration of isochaihulactone （10 mg.kg^-1·d^-1, sc） for 7 weeks concomitant with D-gal injection significantly increased superoxide dismutase and glutathione peroxidase activities and decreased the MDA level in plasma. Furthermore, H＆E staining to quantify cell death within hippocampus showed that percentage of pyknotic nuclei in the D-gal-treated mice were much higher than in control. Conclusion： The results suggest that isochaihulactone exerts potent anti-aging effects against D-gal in mice possibly via antioxidative mechanisms.

Obesity is a major metabolic disorder driven by excessive adipogenesis and lipid accumulation. This study investigated the anti-obesity effects and molecular mechanisms of Antrodia cinnamomea alcohol extract (ACE) in 3T3-L1 preadipocytes and a high-fat diet (HFD)-induced obesity mouse model. In vitro, Antrodia cinnamomea alcohol extract significantly inhibited adipocyte differentiation and lipid accumulation in 3T3-L1 cells by downregulating PPARγ and C/EBPα, while activating the AMPK pathway and suppressing MAPK signaling. In vivo, Antrodia cinnamomea alcohol extract administration reduced body weight, adipose tissue mass, and liver lipid accumulation in high-fat diet-fed mice, ameliorating non-alcoholic fatty liver disease (NAFLD) symptoms. Transcriptomic analysis of adipose tissue revealed that Antrodia cinnamomea alcohol extract modulated key gene expression profiles related to fatty acid metabolism and adipogenesis, suppressing lipid synthesis while enhancing β-oxidation. Furthermore, Antrodia cinnamomea alcohol extract rebalanced gut microbiota, increasing beneficial bacterial populations such as Akkermansia and Bifidobacterium, while reducing pro-inflammatory Escherichia-Shigella species. These findings demonstrate that Antrodia cinnamomea alcohol extract exerts multifaceted anti-obesity effects by regulating lipid metabolism, adipogenesis pathways, and gut microbiota composition, highlighting its potential as a natural therapeutic agent for obesity management.

Bone tissue engineering has seen significant advancements with innovative scaffold fabrication techniques such as 3D printing. This review focuses on enhancing polycaprolactone (PCL) scaffold properties through structural modifications, including surface treatments, pore architecture adjustments, and the incorporation of biomaterials like hydroxyapatite (HA). These modifications aim to improve scaffold conformation, cellular behavior, and mechanical performance, with particular emphasis on the role of mesenchymal stem cells (MSCs) in bone regeneration. The review also explores the potential of integrating nanomaterials and graphene oxide (GO) to further enhance the mechanical and biological properties of PCL scaffolds. Future directions involve optimizing scaffold structures and compositions for improved bone tissue regeneration outcomes.

To explore how the immune system controls clearance of SARS-CoV-2, we used a single-cell, mass cytometry-based proteomics platform to profile the immune systems of 21 patients who had recovered from SARS-CoV-2 infection without need for admission to an intensive care unit or for mechanical ventilation. We focused on receptors involved in interactions between immune cells and virus-infected cells. We found that the diversity of receptor repertoires on natural killer (NK) cells was negatively correlated with the viral clearance rate. In addition, NK subsets expressing the receptor DNAM1 were increased in patients who more rapidly recovered from infection. Ex vivo functional studies revealed that NK subpopulations with high DNAM1 expression had cytolytic activities in response to target cell stimulation. We also found that SARS-CoV-2 infection induced the expression of CD155 and nectin-4, ligands of DNAM1 and its paired coinhibitory receptor TIGIT, which counterbalanced the cytolytic activities of NK cells. Collectively, our results link the cytolytic immune responses of NK cells to the clearance of SARS-CoV-2 and show that the DNAM1 pathway modulates host-pathogen interactions during SARS-CoV-2 infection.

Astragaloside II (AS II) extracted from Astragalus membranaceus has been reported to promote tissue wound repair. However, the effect of AS II on inflammatory bowel disease is unknown. We investigated the effects and mechanism of AS II on intestinal wound healing in both in vitro and in vivo models. Human intestinal Caco-2 cells were treated with multiple concentrations of AS II to assess cell proliferation, scratch wound closure, L-arginine uptake, cationic amino acid transporter activity, and activation of the mTOR signaling pathway. These effects were also measured in a mouse model of colitis. AS II promoted wound closure and increased cell proliferation, L-arginine uptake, CAT1 and CAT2 protein levels, total protein synthesis, and phosphorylation of mTOR, S6K, and 4E-BP1 in Caco-2 cells. These effects were suppressed by lysine or rapamycin treatment, suggesting that the enhanced arginine uptake mediates AS II-induced wound healing. Similar results were also observed in vivo . Our findings indicate that AS II can contribute to epithelial barrier repair following intestinal injury, and may offer a therapeutic avenue in treating irritable bowel disease.

Recent discoveries of the photoresponse of molybdenum disulfide (MoS 2 ) have shown the considerable potential of these two-dimensional transition metal dichalcogenides for optoelectronic applications. Among the various types of photoresponses of MoS 2 , persistent photoconductivity (PPC) at different levels has been reported. However, a detailed study of the PPC effect and its mechanism in MoS 2 is still not available, despite the importance of this effect on the photoresponse of the material. Here, we present a systematic study of the PPC effect in monolayer MoS 2 and conclude that the effect can be attributed to random localized potential fluctuations in the devices. Notably, the potential fluctuations originate from extrinsic sources based on the substrate effect of the PPC. Moreover, we point out a correlation between the PPC effect in MoS 2 and the percolation transport behavior of MoS 2 . We demonstrate a unique and efficient means of controlling the PPC effect in monolayer MoS 2 , which may offer novel functionalities for MoS 2 -based optoelectronic applications in the future.