Identification of a prognostic signature based on five ferroptosis-related genes for diffuse large B-cell lymphoma.

BACKGROUND: Therapies for diffuse large B-cell lymphoma (DLBCL) are limited due to the diverse gene expression profiles and complicated immune microenvironments, making it an aggressive lymphoma. Beyond this, researches have shown that ferroptosis contributes to tumorigenesis, progression, and metastasis. We thus are interested to dissect the connection between ferroptosis and disease status of DLBCL. We aim at generating a valuable prognosis gene signature for predicting the status of patients of DLBCL, with focus on ferroptosis-related genes (FRGs). OBJECTIVE: To examine the connection between ferroptosis-related genes (FRGs) and clinical outcomes in DLBCL patients based on public datasets. METHODS: An expression profile dataset for DLBCL was downloaded from GSE32918 (https://www.ncbi.nlm.nih.gov/geo/ query/acc.cgi?acc=gse32918), and a ferroptosis-related gene cluster was obtained from the FerrDb database (http://www. zhounan.org/ferrdb/). A prognostic signature was developed from this gene cluster by applying a least absolute shrinkage and selection operator (LASSO) Cox regression analysis to GSE32918, followed by external validation. Its effectiveness as a biomarker and the prognostic value was determined by a receiver operator characteristic curve mono factor analysis. Finally, functional enrichment was evaluated by the package Cluster Profiler of R. RESULTS: Five ferroptosis-related genes (FRGs) (GOP1, GPX2, SLC7A5, ATF4, and CXCL2) associated with DLBCL were obtained by a multivariate analysis. The prognostic power of these five FRGs was verified by TCGA (https://xenabrowser.net/datapages/?dataset=TCGA.DLBC.sampleMap%2FHiSeqV2_PANCAN&host=https%3A%2F%2Ftcga.xenahubs.net&removeHub=https%3A%2F%2Fxena.treehouse.gi.ucsc.edu%3A44) and GEO (https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=gse 32918) datasets, with ROC analyses. KEGG and GO analyses revealed that upregulated genes in the high-risk group based on the gene signature were enriched in receptor interactions and other cancer-related pathways, including pathways related to abnormal metabolism and cell differentiation. CONCLUSION: The newly developed signature involving GOP1, GPX2, SLC7A5, ATF4, and CXCL2 has the potential to serve as a prognostic biomarker. Furthermore, our results provide additional support for the contribution of ferroptosis to DLBCL.

Incidence and survival of second primary non-Hodgkin lymphoma: A Surveillance, Epidemiology, and End Results-based cohort study.

BACKGROUND: The aim of this study was to investigate patient survival and factors associated with survival in second primary non-Hodgkin lymphoma (NHL) compared with the first primary NHL. METHODS: The retrospective cohort study used data from the Surveillance, Epidemiology, and End Results (SEER) database between 2000 and 2014. Demographic characteristics, histological types, Ann Arbor stage, and treatment information were collected. Cox proportional hazard models were used to estimate hazard ratios (HRs) and 95% confidence intervals (CIs) for factors associated with overall survival (OS) and cancer-specific survival (CSS) in the first and second primary NHLs. RESULTS: Of 318,168 cases followed for 5 years, 299,248 patients developed the first primary NHL and 18,920 patients developed the second primary NHL. This study identified a rising incidence of first and second primary NHL from 2000 to 2014. For the second primary NHL, the OS risk was higher when compared to the first primary NHL (HR: 1.13, 95% CI: 1.11 to 1.15, P <0.001). Risk factors that negatively affected OS in the first primary NHL included being male, over 40 years of age, certain marital statuses, specific histological types, and advanced disease stages. In contrast, being of White race and having histological types such as Follicular Lymphoma (FL), Marginal Zone Lymphoma (MZL), and mantle B-cell NHL were associated with better OS outcomes. Treatments like surgery, radiation therapy, and chemotherapy were associated with a lower risk of OS and CSS in the first primary NHL. For the second primary NHL, the detrimental risk factors were similar but also included being over the age of 60. Certain histological types showed a lower OS risk relative to diffuse Large B-cell Lymphoma (DLBCL). While surgery and chemotherapy were beneficial for OS, radiation therapy did not improve survival in second primary NHL cases. Notably, undergoing chemotherapy for the first primary cancer increased the OS risk in the second primary NHL, whereas surgery and radiation seemed to offer a protective effect against OS risk in the second primary NHL (all P <0.05). CONCLUSION: Our findings emphasize the need for tailored strategies in managing the second primary NHL, given the distinct survival patterns and risk factor profiles compared to the first primary NHL. Future research should aim to further elucidate these differences to improve prognosis and treatment approaches for second primary NHL patients.

Study on the treatment of postmenopausal osteoporosis with quercetin in Liuwei Dihuang Pill based on network pharmacology.

BACKGROUND: Liuwei Dihuang Pill (LP) was verified to alleviate postmenopausal osteoporosis (PMOP) development. Nevertheless, the major constituent of LP and the related network pharmacology study remain unexplored. METHODS: Protein-protein interaction was established to identify the downstream target of LP in PMOP, and the related signaling pathway was investigated by bioinformatics analysis. MC3T3-E1 cells were added to ferric ammonium citrate (FAC) to mimic osteoporosis in vitro. The osteoblasts were identified by Alizarin red staining. Western blot was applied to evaluate protein levels. In addition, Cell Counting Kit-8 (CCK8) assay was applied to assess cell viability, and cell apoptosis was assessed by flow cytometry. RESULTS: Quercetin was the major constituent of LP. In addition, quercetin significantly reversed FAC-induced inhibition of osteogenic differentiation in MC3T3-E1 cells. In addition, quercetin notably abolished the FAC-induced upregulation of Bax, Caspase-3, FOS, JUN, TGFB1 and PPARD. In contrast, Bcl-2, p-mTOR/mTOR, p-AKT/AKT and p-PI3K/PI3K levels in MC3T3-E1 cells were reduced by FAC, which was restored by quercetin. Meanwhile, FAC notably inhibited the viability of MC3T3-E1 cells via inducing apoptosis, but this impact was abolished by quercetin. Furthermore, quercetin could reverse pcDNA3.1-FOS-mediated growth of FAC-treated osteoblasts by mediating PI3K/AKT/mTOR signaling. CONCLUSION: Quercetin alleviated the progression of PMOP via activation of PI3K/AKT/mTOR signaling. Hence, this study would shed novel insights into discovering new methods against PMOP.

Prevalence, emotional and follow-up burden of insulin injection-related needle-stick injuries among clinical nurses in Shaanxi Province, west of China: A cross-sectional study.

AIMS AND OBJECTIVES: The aim of this study was to investigate the prevalence, emotional and follow-up burden of insulin injection-related needle-stick injuries among clinical nurses. BACKGROUND: needle-stick injures introduce statistically significant occupational hazards to healthcare workers. Although the large proportion of the needles injuries attributed to insulin injection, research evidence about the prevalence, emotional and follow-up burden of such injures is lacking. DESIGN: Cross-sectional study. METHODS: 5389 nurses were recruited from 45 hospitals in Shaanxi, China, from November 2018 to July 2019. Participants were administrated with a questionnaire specifically developed for this study. Descriptive statistics were used to present the findings. RESULTS: All 5,389 nurses responded to the survey, of which 396 (7.4%) participants experienced 620 insulin injection-related needle-stick injuries in the past year, representing an annual prevalence of 115.0 per 1000 nurses. The annual prevalence of infection caused by the injuries was 18.7 per 1000 nurses. The injuries occurred most frequently when nurses were recapping the needle (42.4%). In the majority (98.4%) of the injuries, the hurt nurses took proper immediate actions. However, only 30.3% of nurses reported the injuries to the administrative staff, and in 43.2% of the injuries, the nurses refused or discontinued the suggested follow-up. A large proportion (58.6%) of the hurt nurses experienced emotional changes. Multivariate logistic regression showed that department, removing and/or setting back needle caps with bare hands, frequency of insulin pen and syringes are associated with the incidence of insulin injection-related needle-stick injuries. This paper is reported following the STROBE recommendations. CONCLUSIONS: This survey demonstrated a considerably high prevalence of insulin injection-related needle-stick injuries among clinical nurses. Even though the majority of the hurt nurses took proper immediate actions, a large quantity of them failed to report the accidents to the administrative staff and complete the suggested follow-up. Nurses who suffered from insulin injection-related needle-stick injuries were subject to various negative emotional changes. It portends a statistically significant risk to occupational health management for nurses. RELEVANCE TO CLINICAL PRACTICE: Scientific preventive and management strategies are desirable in order to minimize the consequences of insulin injection-related needle-stick injuries.

MYCN contributes to the sensitization of acute myelogenous leukemia cells to cisplatin by targeting SRY-box transcription factor 4.

Little was known about the role of MYCN in drug resistance in acute myeloid leukemia (AML). We investigated impacts of MYCN on sensitization of AML cell to cisplatin, and its interaction with SRY-box transcription factor 4 (SOX4). In vitro, human AML cell lines HL60 and KG-1 were transfected with the overexpression plasmids and specific small interfering RNA of MYCN (siMYCN), or siSOX4, and then treated with 2 µg/mL cisplatin. MTT and flow cytometry assays were performed to estimate the viability and apoptosis of AML cell. The expressions of MYCN and SOX4 in AML patients and the associations between these two were analyzed by bioinformatics analysis, luciferase assay and quantitative reverse transcription polymerase chain reaction (qRT-PCR). In vivo, transfected HL60 cells were injected into BALB/c nude mice. The tumor size and weight were recorded, and the expressions of MYCN and SOX4 were determined with immunohistochemistry. MYCY and SOX4 were highly-expressed in AML, and their expressions were positively correlated. In AML cells, the treatment using cisplatin induced higher MYCN expression and augmented apoptosis, whereas decreasing the viability. Silencing of MYCN enhanced the tumor-suppressive effects of cisplatin on AML by decreasing cell viability and increasing apoptosis. The promoter of SOX4 was targeted by MYCN. Lower cell viability and higher apoptosis rate were seen in AML cells transfected with siSOX4, the effects of which were partially reversed by overexpressed MYCN. Silencing of MYCN retarded the tumor growth and SOX4 expression in tumor in vivo. Collectively, MYCN sensitized AML cells to cisplatin by targeting SOX4.

Education level as a predictor of survival in patients with multiple myeloma.

BACKGROUND: Disparities in multiple myeloma (MM) prognosis based on sociodemographic factors may exist. We investigated whether education level at diagnosis influenced Chinese MM patient outcomes. METHODS: We performed a multicenter retrospective analysis of data from 773 MM patients across 9 centers in China from 2006 to 2019. Sociodemographic and clinical factors at diagnosis and treatment regimens were recorded, and univariate and multivariate analyses were performed. RESULTS: Overall, 69.2% of patients had low education levels. Patients with low education levels differed from those with high education levels in that they were more likely to be older, and a higher proportion lived in rural areas, were unemployed, had lower annual incomes and lacked insurance. Additionally, compared to patients with high education levels, patients with low education levels had a higher proportion of international staging system (ISS) stage III classification and elevated lactate dehydrogenase (LDH) levels and underwent transplantation less often. Patients with high education levels had a median progression-free survival (PFS) of 67.50 (95% confidence interval (CI): 51.66-83.39) months, which was better than that of patients with low education levels (30.60 months, 95% CI: 27.38-33.82, p < 0.001). Similarly, patients with high education levels had a median overall survival (OS) of 122.27 (95% CI: 117.05-127.49) months, which was also better than that of patients with low education levels (58.83 months, 95% CI: 48.87-62.79, p < 0.001). In the multivariable analysis, patients with high education levels had lower relapse rates and higher survival rates than did those with low education level in terms of PFS and OS (hazard ratio (HR) = 0.50 [95% CI: 0.34-0.72], p < 0.001; HR = 0.32 [0.19-0.56], p < 0.001, respectively). CONCLUSIONS: Low education levels may independently predict poor survival in MM patients in China.

Development of AAV Variants with Human Hepatocyte Tropism and Neutralizing Antibody Escape Capacity.

Adeno-associated virus (AAV) vectors have been successfully used in patients with bleeding disorders and blindness. For human liver targeting, two major factors restrict effective AAV transduction after systemic administration of AAV vectors: human hepatocyte tropism and neutralizing antibodies (Nabs). In this study, we attempted to isolate AAV variants with the ability to transduce human hepatocytes and escape Nabs using a directed evolution approach in vivo. After four cycles of selection, 14 AAV capsid mutants were identified from a capsid shuffling library selected in the presence of human Intravenous Immunoglobulin (IVIG) and isolated from human hepatocytes xenografted into chimeric mice. AAV neutralization assays using IVIG showed that most of the mutants showed the Nab escape pattern in a manner similar to that of AAV8 or AAV9 and better than that of other AAV serotypes. Different mutants displayed varying capacities to escape Nab activity from individual serum samples collected from healthy subjects or hemophilia patients. The mutant AAV LP2-10 was found in 12 colonies out of 25, which was composed of capsids from AAV serotypes 2, 6, 8, and 9, with VP3 subunits derived from AAV8 swapped with AAV6 from residues 261 to 272. The mutant AAV LP2-10 manifested a higher ability than that of other serotypes to escape Nabs in IVIG and most human serum samples. After injection of AAV vectors encoding a self-complementary GFP cassette into chimeric mice, LP2-10 transduced human hepatocytes with efficiency similar to that of AAV8. In summary, AAV mutants can be isolated in humanized mice with both human hepatocyte tropism and the ability to evade Nab activity.

The association between ambient temperature and clinical visits for inflammation-related diseases in rural areas in China.

BACKGROUND: The association between temperature and mortality has been widely reported. However, it remains largely unclear whether inflammation-related diseases, caused by excessive or inappropriate inflammatory reaction, may be affected by ambient temperature, particularly in low-income areas. OBJECTIVES: To explore the association between ambient temperature and clinical visits for inflammation-related diseases in rural villages in the Ningxia Hui Autonomous Region, China, during 2012─2015. METHODS: Daily data on inflammation-related diseases and weather conditions were collected from 258 villages in Haiyuan (161 villages) and Yanchi (97 villages) counties during 2012─2015. A Quasi-Poisson regression with distributed lag non-linear model was used to examine the association between temperature and clinical visits for inflammation-related diseases. Stratified analyses were performed by types of diseases including arthritis, gastroenteritis, and gynecological inflammations. RESULTS: During the study period, there were 724,788 and 288,965 clinical visits for inflammation-related diseases in Haiyuan and Yanchi, respectively. Both exposure to low (RR: 2.045, 95% CI: 1.690, 2.474) and high temperatures (RR: 1.244, 95% CI: 1.107, 1.399) were associated with increased risk of total inflammation-related visits in Haiyuan county. Low temperatures were associated with increased risks of all types of inflammation-related diseases in Yanchi county (RR: 4.344, 95% CI: 2.887, 6.535), while high temperatures only affected gastroenteritis (RR: 1.274, 95% CI: 1.040, 1.561). Moderate temperatures explained approximately 26% and 33% of clinical visits due to inflammation-related diseases in Haiyuan and Yanchi, respectively, with the burden attributable to cold exposure higher than hot exposure. The reference temperature values ranged from 17 to 19 in Haiyuan, and 12 to 14 in Yanchi for all types of clinical visits. CONCLUSIONS: Our findings add additional evidence for the adverse effect of suboptimal ambient temperature and provide useful information for public health programs targeting people living in rural villages.

Short hairpin RNA attenuates liver fibrosis by regulating the PPAR­Î³ and NF­κB pathways in HBV­induced liver fibrosis in mice.

Progressive liver fibrosis, caused by chronic viral infection and metabolic disorders, results in the development of cirrhosis and hepatocellular carcinoma. However, no antifibrotic therapies have been approved to date. In our previous study, adeno­associated virus (AAV) short hairpin RNAs (shRNAs) targeting hepatitis B virus (HBV) and transforming growth factor (TGF)­ß administration could persistently inhibit HBV replication and concomitantly prevent liver fibrosis. However, the differentially expressed proteins and critical regulatory networks of AAV­shRNA treatment remain unclear. Accordingly, in the present study, we aimed to analyze differentially expressed proteins in the liver of AAV­shRNA­treated mice with HBV infection and liver fibrosis using isobaric tags for relative and absolute quantitation (iTRAQ)­based quantitative proteomics and to elucidate the underlying antifibrotic mechanisms. In total 2,743 proteins were recognized by iTRAQ­based quantitative proteomics analysis. Gene Ontology analysis revealed that the differentially expressed proteins mostly participated in peptide metabolism in the biological process category, cytosolic ribosomes in the cell component category, and structural constituents of ribosomes in the molecular function category. Kyoto Encyclopedia of Genes and Genomes pathway analysis indicated that oxidative stress and the peroxisome proliferator­activated receptor (PPAR) signaling pathway were activated after treatment. Verification studies revealed that AAV­shRNAs inhibited hepatic stellate cell activation and inflammation by suppressing nuclear factor­κB p65 phosphorylation and α­smooth muscle actin expression via upregulation of PPAR­Î³. Hepatocytes steatosis was also decreased by activating the PPAR signaling pathway and improving lipid metabolism. The expression level of TGF­ß was decreased due to upregulation of PPAR­Î³ expression and direct inhibition using AAV­shRNA targeting TGF­ß. TGF­ß­induced oxidative stress was suppressed by increasing glutathione S­transferase Pi 1 and reducing peroxiredoxin 1. Collectively, the present results indicated that AAV­shRNAs were effective in modulating liver fibrosis by reducing oxidative stress, inflammation and activating the PPAR signaling pathway.

Epidemiological and clinical characteristics of Dengue virus outbreaks in two regions of China, 2014 - 2015.

Dengue virus (DENV), a single-stranded RNA virus and Flaviviridae family member, is transmitted by Aedes aegypti and Aedes albopictus mosquitoes. DENV causes dengue fever, which may progress to severe dengue. Hospital-based surveillance was performed in two Chinese regions, Guangzhou and Xishuangbanna, during the dengue epidemics in 2014 and 2015, respectively. Acute-phase serum was obtained from 133 patients with suspected dengue infections during the peak season for dengue cases. Viremia levels, virus sero-positivity, serotype distribution, infection type, clinical manifestations and virus phylogenetics were investigated. Of the 112 DENV-confirmed cases, 92(82.14%) were IgM antibody-positive for DENV, and 69(51.88%) were positive for DENV RNA. From these cases, 47(41.96%) were classified as primary infections, 39(34.82%) as secondary infections and 26 (23.21%) as undetermined infections. The viremia levels were negatively correlated with IgM presence, but had no relationship with the infection type. DENV-1 genotype V dominated in Guangzhou, whereas the DENV-2 Cosmopolitan genotype dominated in Xishuangbanna, where fewer DENV-1 genotype I cases occurred. DENV-2 is associated with severe dengue illness with more serious clinical issues. The strains isolated during 2014-2015 are closely related to the isolates obtained from other Chinese regions and to those isolated recently in Southeast Asian countries. Our results indicate that DENV is no longer an imported virus and is now endemic in China. An extensive seroepidemiological study of DENV and the implementation of vector control measures against it are now warranted in China.

Author Correction: Enhanced antiviral and antifibrotic effects of short hairpin RNAs targeting HBV and TGF-ß in HBV-persistent mice.

A correction to this article has been published and is linked from the HTML and PDF versions of this paper. The error has been fixed in the paper.

Impact of ambient temperature on clinical visits for cardio-respiratory diseases in rural villages in northwest China.

BACKGROUND: The association between temperature and cardio-respiratory disease in urban areas has been widely reported but there is limited information from populations living in rural areas that may be disproportionately affected by climate change. OBJECTIVES: To quantify the associations between daily temperature and clinical visits due to cardiovascular and/or respiratory disease in rural villages in the Ningxia Hui Autonomous Region, China over 2012-2015. METHODS: Daily data on clinical visits and weather conditions were collated from 203 villages. A quasi-Poisson regression with distributed lag non-linear model was used to examine the associations between daily temperature and clinical visits up to 28days, after controlling for potential confounders. RESULTS: Over three years, 158,733 and 1,272,212 clinical visits were recorded for cardiovascular and respiratory diseases, respectively. Both low and high temperatures were associated with an increased risk of clinical visits for cardiovascular-related conditions, whereas only low temperatures were associated with increased clinical visits related to respiratory illness. The cold effect on cardiovascular visits appeared at the lag 6th day and persisted until the 22nd day, resulting in a cumulative relative risk (RR) 1.55 (95% CI: 1.26-1.92), compared with the minimum-clinical visit temperature. The cold effect on respiratory visits appeared immediately and lasted over the lag 0-28days, with a cumulative RR 2.96 (2.74-3.21). Suboptimal temperature accounted for approximately 13% and 26% of clinic visits due to cardiovascular and respiratory disorders, respectively, with the majority of cases attributable to moderate - rather than extreme - cold temperature. CONCLUSIONS: In rural settings, sub-optimal temperatures explained nearly one quarter of all clinical visits due to cardiovascular and respiratory diseases. Although extreme cold temperature had a stronger, more immediate, prolonged effect on respiratory disease than for cardiovascular disease, moderately cold temperatures accounted for most of the overall burden of clinical visits.

MiR-199a-3p suppresses proliferation and invasion of prostate cancer cells by targeting Smad1.

OBJECTIVES: This study was intended to analyze effects of miR-199a-3p and Smad1 on proliferation, migration and invasion of prostate cancer (PCa) cells. RESULTS: MiR-199a-3p was significantly decreased in PCa tissues in comparison to that in adjacent normal tissues (P < 0.05). Over-expressed miR-199a-3p markedly suppressed proliferation and invasion of PCa cells (P < 0.05). MiR-199a-3p was negatively correlated with Smad1 expression, and overexpression of Smad1 could antagonize the effects of miR-199a-3p on PCa cells. MATERIALS AND METHODS: The PCa tissues and their adjacent normal tissues were collected from 54 PCa patients. Expressions of miR-199a-3p and Smad1 mRNA in tissues and cells were evaluated with real-time quantitative polymerase chain reaction (RT-qPCR), and immunohistochemistry assay was used to detect Smad1 protein expressions. The target relationship between miR-199a-3p and Smad1 was assessed by luciferase reporter assay. The PCa cell lines (i.e. PC-3 cells) were transfected with miR-199a-3p mimics and Smad1-cDNA. MTT and Transwell assays were applied to detect proliferative, migratory and invasive abilities of PCa cells. CONCLUSIONS: MiR-199a-3p suppressed proliferation and invasion of PCa cells by targeting Smad1.

High concentration of miR-133 is a useful marker for the diagnosis of lymphoma- associated hemophagocytic syndrome.

BACKGROUND: Lymphoma associated hemophagocytic syndrome (LAHS) is one of the major adult secondary hemophagocytic lymphohistiocytosis (HLH). Early diagnosis and treatment contribute to improved outcome. No enlarge lymph nodes can often delay the diagnosis of underlying lymphoma. OBJECTIVE: To find out criteria distinguishing LAHS from HLH induced by benign diseases. METHODS: clinical characteristic and laboratory feature of 31 patients with HLH (10 benign disease-associated HLH and 21 LAHS) were analyzed retrospectively. RESULTS: No significantly differences were observed in the levels of LDH, IL-6, IL-10, TNF-α; however, the level of CRP (C reactive protein) and the mean level of sIL-R (soluble interleukin-2 receptor) were higher in patients with LAHS than those with benign disease associated disease associated HLH while ferritin levels were higher in benign disease associated HLH than in LAHS. Consequently, the serum sIL-2R/ferritin ratio of patients with LAHS was markedly higher than that of patients with benign disease associated HLH (0.33 ± 0.23 vs 5.82 ± 3.26, P= 0.0001). In addition, we found out that the mean level of miR-133 (microRNA-133) was significant higher in LAHS than in benign disease associated HLH (18.83 ± 10.44 vs 5.82 ± 3.26, P⩽ 0.0001). CONCLUSION: Serum miR-133 is a new very useful marker for diagnosing of LAHS, but it need further confirmation by further clinical studies.

Proteomic and transcriptomic studies of HBV-associated liver fibrosis of an AAV-HBV-infected mouse model.

BACKGROUND: Human hepatitis B virus (HBV) infection is an important public health issue in the Asia-Pacific region and is associated with chronic hepatitis, liver fibrosis, cirrhosis and even liver cancer. However, the underlying mechanisms of HBV-associated liver fibrosis remain incompletely understood. RESULTS: In the present study, proteomic and transcriptomic approaches as well as biological network analyses were performed to investigate the differentially expressed molecular signature and key regulatory networks that were associated with HBV-mediated liver fibrosis. RNA sequencing and 2DE-MALDI-TOF/TOF were performed on liver tissue samples obtained from HBV-infected C57BL/6 mouse generated via AAV8-HBV virus. The results showed that 322 genes and 173 proteins were differentially expressed, and 28 HBV-specific proteins were identified by comprehensive proteomic and transcriptomic analysis. GO analysis indicated that the differentially expressed proteins were predominantly involved in oxidative stress, which plays a key role in HBV-related liver fibrosis. Importantly, CAT, PRDX1, GSTP1, NXN and BLVRB were shown to be associated with oxidative stress among the differentially expressed proteins. The most striking results were validated by Western blot and RT-qPCR. The RIG-I like receptor signaling pathway was found to be the major signal pathway that changed during HBV-related fibrosis. CONCLUSIONS: This study provides novel insights into HBV-associated liver fibrosis and reveals the significant role of oxidative stress in liver fibrosis. Furthermore, CAT, BLVRB, NXN, PRDX1, and IDH1 may be candidates for detection of liver fibrosis or therapeutic targets for the treatment of liver fibrosis.

Enhanced antiviral and antifibrotic effects of short hairpin RNAs targeting HBV and TGF-ß in HBV-persistent mice.

The hepatitis B virus (HBV) causes acute and chronic liver infection, which may lead to liver cirrhosis and hepatocellular carcinoma. Current treatments including interferons and nucleotide analogs, have limited therapeutic effects, underscoring the need to identify effective therapeutic options to inhibit HBV replication and prevent complications. Previous animal models mimicking chronic HBV infection do not faithfully reflect disease progression in humans. Here, we used our established HBV-persistent mouse line with liver fibrosis to evaluate the efficacy of novel therapies. The combination of two short hairpin RNAs (dual-shRNA) against different coding regions of HBV delivered by a self-complementary AAV vector showed better antiviral effects than single shRNA both in vitro and in HBV-persistent mice. The dual-shRNA also exhibited stronger antifibrotic activity in vivo. Vector carrying shRNA against TGF-ß, though did not inhibit HBV replication alone, enhanced the antiviral and antifibrotic activities of single and dual HBV shRNAs. Co-administration of TGF-ß shRNA and HBV dual-shRNA decreased HBV DNA, HBV RNA, HBsAg, HBeAg, and liver fibrosis markers in serum and tissues, and improved liver morphology more effectively than single treatments. Our results suggest that the combination of shRNAs against HBV and TGF-ß could be developed into a viable treatment for human HBV infection.

Peiminine inhibits colorectal cancer cell proliferation by inducing apoptosis and autophagy and modulating key metabolic pathways.

Peiminine, a compound extracted from the bulbs of Fritillaria thunbergii and traditionally used as a medication in China and other Asian countries, was reported to inhibit colorectal cancer cell proliferation and tumor growth by inducing autophagic cell death. However, its mechanism of anticancer action is not well understood, especially at the metabolic level, which was thought to primarily account for peiminine's efficacy against cancer. Using an established metabolomic profiling platform combining ultra-performance liquid chromatography/tandem mass spectrometry with gas chromatography/mass spectrometry, we identified metabolic alterations in colorectal cancer cell line HCT-116 after peiminine treatment. Among the identified 236 metabolites, the levels of 57 of them were significantly (p < 0.05) different between peiminine-treated and -untreated cells in which 45 metabolites were increased and the other 12 metabolites were decreased. Several of the affected metabolites, including glucose, glutamine, oleate (18:1n9), and lignocerate (24:0), may be involved in regulation of the phosphoinositide 3-kinase/Akt/mammalian target of rapamycin (mTOR) pathway and in the oxidative stress response upon peiminine exposure. Peiminine predominantly modulated the pathways responsible for metabolism of amino acids, carbohydrates, and lipids. Collectively, these results provide new insights into the mechanisms by which peiminine modulates metabolic pathways to inhibit colorectal cancer cell growth, supporting further exploration of peiminine as a potential new strategy for treating colorectal cancer.

Development of Patient-specific AAV Vectors After Neutralizing Antibody Selection for Enhanced Muscle Gene Transfer.

A major hindrance in gene therapy trials with adeno-associated virus (AAV) vectors is the presence of neutralizing antibodies (NAbs) that inhibit AAV transduction. In this study, we used directed evolution techniques in vitro and in mouse muscle to select novel NAb escape AAV chimeric capsid mutants in the presence of individual patient serum. AAV mutants isolated in vitro escaped broad patient-specific NAb activity but had poor transduction ability in vivo. AAV mutants isolated in vivo had enhanced NAb evasion from cognate serum and had high muscle transduction ability. More importantly, structural modeling identified a 100 amino acid motif from AAV6 in variable region (VR) III that confers this enhanced muscle tropism. In addition, a predominantly AAV8 capsid beta barrel template with a specific preference for AAV1/AAV9 in VR VII located at threefold symmetry axis facilitates NAb escape. Our data strongly support that chimeric AAV capsids composed of modular and nonoverlapping domains from various serotypes are capable of evading patient-specific NAbs and have enhanced muscle transduction.