Organohalogen contaminants threaten the survival of indo-pacific humpback dolphin calves in their largest habitat.

As long-lived apex predators, marine mammal adults often accumulate alarmingly levels of environmental contaminants. Nevertheless, the accumulation and risks of these contaminants in the critical calf stage of marine mammals remain largely unknown. Here, we investigated the exposure status and health risks of 74 organohalogen contaminants (OHCs) in Indo-Pacific humpback dolphin calves (Sousa chinensis) collected from the Pearl River Estuary (PRE), China, during 2005-2019. Our findings revealed moderate levels of polychlorinated biphenyls (PCBs), medium-high levels of dichlorodiphenyltrichloroethanes (DDTs) and hexachlorocyclohexanes (HCHs), and the highest levels of polybrominated diphenyl ethers (PBDEs) and alternative halogenated flame retardants (AHFRs) compared to those reported for cetaceans elsewhere. Traditional OHCs like DDTs, PCBs, and PBDEs did not exhibit significant decreasing trends in the dolphin calves despite global restrictions on these compounds, and AHFRs as emerging OHCs showed an increasing trend over the study period. Risk quotients of DDTs, HCHs, PBDEs, and PCBs in most of the dolphin samples were > 1, indicating that humpback dolphin calves may have suffered long-term threats from OHC exposure. The significant correlation observed between the traditional OHC levels and the stranding death number of the dolphin calves suggests these OHCs may impact the survival of this endangered species.

Liver resection versus microwave ablation for solitary and small (≤ 3 cm) HCC with early recurrence in different stages of liver cirrhosis: A propensity score matching study.

BACKGROUND: This study aimed to compare the effectiveness of liver resection (LR) and microwave ablation (MWA) in hepatocellular carcinoma (HCC) patients with early recurrence and varying stages of cirrhosis. METHOD: This study analyzed patients with HCC who underwent hepatectomy and experienced early tumor recurrence (≤3 cm) between December 2002 and December 2020 at the Tongji Hospital. Treatment effectiveness was assessed using a propensity score matching (PSM) analysis. RESULTS: This study included 295 patients (106, LR; 189, MWA), 86 patients in each of the 2 groups were chosen for further comparison, after PSM. After PSM, both LR and MWA demonstrated similar recurrence-free survival (RFS) and overall survival (OS) rates (p = 0.060 and p = 0.118, respectively). However, the LR group had more treatment-related complications. In patients with moderate or severe cirrhosis, no significant differences in RFS or OS rates were found between the LR and MWA groups (p = 0.779 and p = 0.772, respectively). In patients without cirrhosis or with mild cirrhosis, LR showed better RFS and OS rates than MWA (p = 0.024 and p = 0.047, respectively). Multivariate analysis after PSM identified moderate or severe cirrhosis and recurrence intervals ≤12 months as independent predictors of poor RFS and OS in patients with early recurrence of HCC. CONCLUSION: LR is more effective than MWA for early recurrence of HCC in patients without cirrhosis or with mild cirrhosis, showing improved RFS and OS rates. In patients with moderate or severe cirrhosis, the OS and RFS were statistically equal between the two therapies. However, MWA may be preferred owing to its low complication rate.

Bioaccumulation and Potential Endocrine Disruption Risk of Legacy and Emerging Organophosphate Esters in Cetaceans from the Northern South China Sea.

Despite the increasing health risks shown by the continuous detection of organophosphate esters (OPEs) in biota in recent years, information on the occurrence and potential risks of OPEs in marine mammals remains limited. This study conducted the first investigation into the body burdens and potential risks of 10 traditional OPEs (tOPEs) and five emerging OPEs (eOPEs) in 10 cetacean species (n = 84) from the northern South China Sea (NSCS) during 2005-2021. All OPEs, except for 2-ethylhexyl diphenyl phosphate (EHDPHP), were detected in these cetaceans, indicating their widespread occurrence in the NSCS. Although the levels of the ∑10tOPEs in humpback dolphins remained stable from 2005 to 2021, the concentrations of the ∑5eOPEs showed a significant increase, suggesting a growing demand for these new-generation OPEs in South China. Dolphins in proximity to urban regions generally exhibited higher OPE concentrations than those from rural areas, mirroring the environmental trends of OPEs occurring in this area. All OPE congeners, except for EHDPHP, in humpback dolphins exhibited a maternal transfer ratio >1, indicating that the dolphin placenta may not be an efficient barrier for OPEs. The observed significant correlations between levels of OPEs and hormones (triiodothyronine, thyroxine, and testosterone) in humpback dolphins indicated that OPE exposures might have endocrine disruption effects on the dolphin population.

First determination of elevated levels of plastic additives in finless porpoises from the South China Sea.

Plastic additives, such as organophosphate esters (OPEs) and phthalate esters (PAEs), are raising public concerns due to their widespread presence and potential health risks. Nonetheless, the occurrences and potential health risks of these additives in marine mammals remain limited. Here, we first investigated the accumulation patterns and potential risks of OPEs and metabolites of PAEs (mPAEs) in Indo-Pacific finless porpoises inhabiting the northern South China Sea (NSCS) during 2007-2020. The average hepatic concentrations of ∑15OPEs and ∑16mPAEs in the NSCS finless porpoises were 53.9 ± 40.7 and 98.6 ± 54.8 ng/g ww, respectively. The accumulation of mPAEs and OPEs in the finless porpoises is associated with the chemical structures of the compounds. ∑5halogenated-OPEs were the most dominant category (62.6%) of ∑15OPEs, followed by ∑6aryl-OPEs (25.9%) and ∑6nonhalogenated alkyl-OPEs (11.5%). The accumulation of mPAEs displayed a declining trend with increasing alkyl side chain length (C0-C10). Although the hepatic burden of mPAEs in finless porpoises was sex-independent, some OPEs, including TDCIPP, TBOEP, TCIPP, TCrP, TPHP, and TDBPP, exhibited significantly higher concentrations in adult males than in adult females. TDBPP, as a new-generation OPE, exhibited a gradual increase during the study period, suggesting that TDBPP should be prioritized for monitoring in the coastal regions of South China. The estimated hazard quotient indicated that almost all mPAEs and OPEs pose no hazard to finless porpoises, with only DEHP presenting potential health risks to both adult and juvenile finless porpoises.

Prediction of high Ki-67 proliferation index of gastrointestinal stromal tumors based on CT at non-contrast-enhanced and different contrast-enhanced phases.

OBJECTIVES: To evaluate and analyze radiomics models based on non-contrast-enhanced computed tomography (CT) and different phases of contrast-enhanced CT in predicting Ki-67 proliferation index (PI) among patients with pathologically confirmed gastrointestinal stromal tumors (GISTs). METHODS: A total of 383 patients with pathologically proven GIST were divided into a training set (n = 218, vendor 1) and 2 validation sets (n = 96, vendor 2; n = 69, vendors 3-5). Radiomics features extracted from the most recent non-contrast-enhanced and three contrast-enhanced CT scan prior to pathological examination. Random forest models were trained for each phase to predict tumors with high Ki-67 proliferation index (Ki-67>10%) and were evaluated using the area under the receiver operating characteristic curve (AUC) and other metrics on the validation sets. RESULTS: Out of 107 radiomics features extracted from each phase of CT images, four were selected for analysis. The model trained using the non-contrast-enhanced phase achieved an AUC of 0.792 in the training set and 0.822 and 0.711 in the two validation sets, similar to models trained on different contrast-enhanced phases (p > 0.05). Several relevant features, including NGTDM Busyness and tumor size, remained predictive in non-contrast-enhanced and different contrast-enhanced images. CONCLUSION: The results of this study indicate that a radiomics model based on non-contrast-enhanced CT matches that of models based on different phases of contrast-enhanced CT in predicting the Ki-67 PI of GIST. GIST may exhibit similar radiological patterns irrespective of the use of contrast agent, and such radiomics features may help quantify these patterns to predict Ki-67 PI of GISTs. CLINICAL RELEVANCE STATEMENT: GIST may exhibit similar radiomics patterns irrespective of contrast agent; thus, radiomics models based on non-contrast-enhanced CT could be an alternative for risk stratification in GIST patients with contraindication to contrast agent. KEY POINTS: • Performance of radiomics models in predicting Ki-67 proliferation based on different CT phases is evaluated. • Non-contrast-enhanced CT-based radiomics models performed similarly to contrast-enhanced CT in risk stratification in GIST patients. • NGTDM Busyness remains stable to contrast agents in GISTs in radiomics models.

Bile acids regulate SF-1 to alter cholesterol balance in adrenocortical cells via S1PR2.

Glucocorticoid synthesis typically occurs in adrenal cortex and is influenced by cholesterol balance, since cholesterol is the sole precursor of steroids. Bile acids as the signaling molecules, have been shown to promote steroidogenesis in steroidogenic cells. However, whether bile acids directly regulate cholesterol balance remains elusive. In this study, we prepared cholestatic mouse models and cultured human adrenocortical cells (H295R) treated with taurochenodeoxycholic acid (TCDCA) to determine transcription levels of cholesterol metabolism associated genes and cholesterol concentrations in adrenocortical cells. Results showed that common bile duct ligation (CBDL) and chenodeoxycholic acid (CDCA) feeding elevated the mRNA levels of Abca1, Cyp51, Hmgcr, Srb1, and Mc2r in adrenals of mice. Meanwhile, the concentrations of total cholesterol and cholesteryl ester in adrenals of CBDL and CDCA-fed mice were dramatically lowered. The total and phosphorylation levels of HSL in adrenal glands of CBDL mice were also enhanced. Similarly, TCDCA treatment in H295R cells decreased intracellular concentrations of total cholesterol and cholesteryl ester and increased transcription levels of SRB1, MC2R, and HSL as well. Inhibition of bile acids' receptor sphingosine 1-phosphate receptor 2 (S1PR2), extracellular signal-regulated kinase (ERK) phosphorylation, and steroidogenic factor 1 (SF-1) respectively successfully abolished effect of TCDCA on H295R cells. SF-1s was found to be phosphorylated at Thr75 in TCDCA-treated H295R cells. While a mild increase of intracellular cAMP concentration was detected upon TCDCA treatment, inhibition of PKA activity with Rp-Isomer in H295R cells failed to decrease the expression of SF-1 and its target genes. Our findings suggest that conjugated bile acids affect cholesterol balance through regulation of SF-1 in adrenocortical cells so as to provide an adequate cholesterol supply for glucocorticoid synthesis, which improves and enriches our understanding of the mechanism whereby bile acids regulate cholesterol balance to affect adrenal function.

Personalized radiomics signature to screen for KIT-11 mutation genotypes among patients with gastrointestinal stromal tumors: a retrospective multicenter study.

OBJECTIVES: Gastrointestinal stromal tumors (GISTs) carrying different KIT exon 11 (KIT-11) mutations exhibit varying prognoses and responses to Imatinib. Herein, we aimed to determine whether computed tomography (CT) radiomics can accurately stratify KIT-11 mutation genotypes to benefit Imatinib therapy and GISTs monitoring. METHODS: Overall, 1143 GISTs from 3 independent centers were separated into a training cohort (TC) or validation cohort (VC). In addition, the KIT-11 mutation genotype was classified into 4 categories: no KIT-11 mutation (K11-NM), point mutations or duplications (K11-PM/D), KIT-11 557/558 deletions (K11-557/558D), and KIT-11 deletion without codons 557/558 involvement (K11-D). Subsequently, radiomic signatures (RS) were generated based on the arterial phase of contrast CT, which were then developed as KIT-11 mutation predictors using 1408 quantitative image features and LASSO regression analysis, with further evaluation of its predictive capability. RESULTS: The TC AUCs for K11-NM, K11-PM/D, K11-557/558D, and K11-D ranged from 0.848 (95% CI 0.812-0.884), 0.759 (95% CI 0.722-0.797), 0.956 (95% CI 0.938-0.974), and 0.876 (95% CI 0.844-0.908), whereas the VC AUCs ranged from 0.723 (95% CI 0.660-0.786), 0.688 (95% CI 0.643-0.732), 0.870 (95% CI 0.824-0.918), and 0.830 (95% CI 0.780-0.878). Macro-weighted AUCs for the KIT-11 mutant genotype ranged from 0.838 (95% CI 0.820-0.855) in the TC to 0.758 (95% CI 0.758-0.784) in VC. TC had an overall accuracy of 0.694 (95%CI 0.660-0.729) for RS-based predictions of the KIT-11 mutant genotype, whereas VC had an accuracy of 0.637 (95%CI 0.595-0.679). CONCLUSIONS: CT radiomics signature exhibited good predictive performance in estimating the KIT-11 mutation genotype, especially in prediction of K11-557/558D genotype. RS-based classification of K11-NM, K11-557/558D, and K11-D patients may be an indication for choice of Imatinib therapy.

Organotins in fish, shrimp, and cephalopods from the Pearl River Estuary, China: Dietary exposure risk to Indo-Pacific humpback dolphin and human.

Food has regularly been proven to be a key source of exposure to environmental pollutants, drawing attention to the dietary exposure risks of contaminants to mammals with significant daily food intake. Here, the levels of six organotin compounds (OTs) in 18 fish (n = 310), three cephalopods (n = 50), and one shrimp (n = 34) from the Lingdingyang (LDY) and west four region (WFR) of the Pearl River Estuary (PRE) and their dietary exposure risks to Indo-Pacific humpback dolphins and humans were first investigated. Total OT levels ranged from 3.84 to 901. 48 ng/g wet weight (ww) in 22 prey species from the LDY, and from 14.37 to 1364.64 ng/g ww in 19 species from the WFR. The LDY marine species generally accumulated higher butyltin levels but lower phentyltin levels than those in the WFR. All species have a phenyltin degradation index <1 and over 60 % of the sampled species have a butyltin degradation index <1, suggesting the PRE marine species might be exposed to the fresh discharge of OTs. A total of nine marine species exceeded the threshold levels of OT intake for adverse health effects on human juveniles by consumption, all 22 marine species posed high dietary risks to the PRE humpback dolphins. Moreover, probabilistic risk assessment using Monte Carlo simulation revealed that the probabilities of RQ values associated with WFR OT exposure higher than 1 were 18.87 % for human adults, 40.55 % for human juveniles, 100 % for both humpback dolphin adults and humpback dolphin juveniles. Our results highlighted the potentially high dietary exposure risks of OTs to marine mammals and residents in the PRE.

Fatty acids as bioindicators of organohalogen exposure in marine fish from a highly polluted estuary: First insight into small-scale regional differences.

Increasing evidence has revealed the lipid-disrupting effects of organic contaminants on aquatic organisms, raising attention about the efficacy of fatty acids (FAs) as bioindicator of contaminant exposure on marine organisms. Here, we investigated the concentrations of 55 organohalogen contaminants (OHCs), 35 FAs, and their correlations in 15 marine fish species (n = 274) from the estuary outlets of the west four region (WFR) and Lingdingyang (LDY) waters in the Pearl River Estuary (PRE), respectively. Despite the similar OHC profiles, significantly higher concentrations of ∑55OHCs were detected in fish from the LDY than those in the WFR. However, FAs in the LDY fish generally contained lower proportions of polyunsaturated fatty acids than in the WFR fish. A total of 148 and 221 significant correlations between OHCs and FAs were observed in fish samples from the LDY and WFR, respectively, supporting that FAs could be efficient bioindicators of OHC stress in marine fish. However, the low overlaps (14/369) of OHC-FA correlations in fish from the two regions suggested that the bioindicators of OHCs might have spatial heterogeneity. Our results highlighted that FAs likely act as potential bioindicators of OHCs in marine fish, while the regional-specific characteristic of the bioindicators should be considered.

Risk assessment of phthalate metabolites accumulated in fish to the Indo-Pacific humpback dolphins from their largest habitat.

Food has consistently been shown to be an important source of exposure to environmental pollutants, drawing attention to the health risks of pollutants in marine mammals with high daily food intake. Here, the dietary exposure risks posed to the Indo-Pacific humpback dolphins from the Pearl River Estuary (PRE), China, by fourteen phthalate metabolites (mPAEs) were evaluated for the first time. On the basis of liquid chromatography-mass spectrometry (LC-MS/MS) analysis, the levels of ∑14mPAEs in ten main species of prey fish (n = 120) of dolphins ranged from 103.0 to 444.5 ng/g wet weight (ww), among which Bombay duck contained a significantly higher body burden of ∑14mPAEs than other prey species. Phthalic acid (PA), monooctyl phthalate (MnOP), monononyl phthalate (MNP), monoethyl phthalate (MEP), monoethylhexyl phthalate (MEHP), mono (5-carboxy-2-ethylpentyl) phthalate (MECPP), monobutyl phthalate (MBP), and monoisobutyl phthalate (MiBP) all had a trophic magnification factor (TMF) greater than unity, indicating the biomagnification potential of these mPAEs in the marine ecosystem of the PRE. A dietary exposure assessment based on the adjusted reference dose values of phthalates (PAEs) showed that bis (2-ethylhexyl) phthalate (DEHP) and dibutyl phthalate (DBP) may pose a high (HQ > 1) and medium (0.01 < HQ < 1) risk to the dolphin adults and juveniles, respectively. Our results highlight the potential health risks of mPAEs to marine mammals through dietary routes.

Occurrences and potential lipid-disrupting effects of phthalate metabolites in humpback dolphins from the South China Sea.

Phthalate esters (PAEs) are ubiquitous environmental contaminants, arising growing public concern. Nevertheless, information on the exposure and risks of PAEs in wildlife remains limited. Here, we conducted the first investigation of the occurrences, spatiotemporal trends, and potential risks of twelve metabolites of PAEs (mPAEs) in 74 humpback dolphins from the northern South China Sea during 2005-2020. All twelve mPAEs (∑12mPAEs: 9.6-810.7 ng g-1 wet weight) were detected in the dolphin liver, and seven major mPAEs showed increasing trends during the study period, indicating high PAE contamination in the coastal environment of South China. Monoethylhexyl phthalate accounted for over half of the ∑12mPAE concentrations. The accumulation of mPAEs in the dolphins was neither age-dependent nor sex-specific. Compared to parent PAEs, mPAEs generally induced higher agonistic effects on the dolphin peroxisome proliferator-activated receptor alpha/gamma (PPARA/G) as master regulators of lipid homeostasis. Although short-term in vitro assays revealed no significant activation of dolphin PPARA/G by tissue-relevant doses of mPAEs, long-term in vivo evidence (i.e., correlations between hepatic mPAEs and blubber fatty acids) suggested that chronic exposure to mPAEs might have impacted lipid metabolism in the dolphin. This study highlighted the potential health risks of PAE exposure on marine mammals.

MiR-221-3p targets HIPK2 to promote diabetic wound healing.

Diabetic foot ulcer is a severe complication of diabetes and is prone to being a chronic non-healing wound. We previously demonstrated that endothelial progenitor cell-derived exosomes, which contain miR-221-3p, alleviate diabetic ulcers. Here, to explore the mechanisms underlying this wound healing, we investigated the potential angiogenic effects of miR-221-3p in vitro using cultured human umbilical vein endothelial cells (HUVECs) and in vivo using a streptozotocin-induced mouse model of diabetes. We found that miR-221-3p promoted HUVEC viability, migration, and capillary-like tube formation. HUVECs cultured in high glucose showed up-regulated expression of homeodomain-interacting protein kinase 2 (HIPK2), a predicted target of miR-221-3p that may decrease angiogenesis. Knockdown of HIPK2 enhanced high glucose-suppressed HUVEC viability, migration, and tube formation, counteracting the effects of high glucose. Using a dual luciferase reporter assay, we found that HIPK2 was indeed a direct target of miR-221-3p. Subcutaneous injection of miR-221-3p agomir into diabetic mice promoted wound healing and suppressed HIPK2 expression in wound margin tissue. These findings indicate that HIPK2, as a direct target of miR-221-3p, contributes to the regulatory role of miR-221-3p in diabetic wound healing and may be a novel therapeutic target for diabetic foot ulcer.

Microplastics in the endangered Indo-Pacific humpback dolphins (Sousa chinensis) from the Pearl River Estuary, China.

Microplastic pollution is a growing concern worldwide. Despite numerous studies showing the occurrence of microplastics in low-trophic level aquatic organisms, microplastic ingestion and contamination in cetaceans, especially those from Asian waters, has been rarely recorded. Here, we investigated stomach microplastic pollution in twelve Indo-Pacific humpback dolphins stranded along the Pearl River Estuary (PRE), China. We also compared microplastic abundances in dolphins stranded near populated urban areas (ZH, n = 6) with those stranded near rural areas (JM, n = 6). Microplastics were detected in all samples, with abundance ranging widely from 11 to 145 items individual-1 (mean ± SD, 53 ± 35.2). Major microplastics were polypropylene and polyethylene fibers, with the size mostly ranging from 1 to 5 mm and the dominant colors of white or transparent. Humpback dolphins from ZH (73 ± 36.8 items individual-1) exhibited a significantly higher average microplastic abundance than those from JM (33 ± 18.3 items individual-1, p < 0.05). In particular, the highest microplastic concentration was identified in the dolphin (SC-ZH01) stranded near the mouth of the Pearl River, whereas the dolphin (SC-JM04) collected at the rural site contained the lowest concentration of microplastics, suggesting the important influence of land-based human activities on the accumulation of microplastics in the PRE. The identification of varied microplastic polymers indicated their complex source scenarios. This study suggests that, as one of top predators in the potential microplastic food chains, this cetacean species could likely serve as an endpoint biomonitoring species of microplastic pollution in the PRE or other similar estuarine ecosystems. Our results highlight the need for more studies towards better understanding the potential impacts of microplastics on this endangered species.

Enhancement and mechanism of vermiculite thermal expansion modified by sodium ions.

Aimed at improving vermiculite's thermal expansibility, a novel method of Na+ modification has been proposed. The modification effects were characterized via X-ray fluorescence spectroscopy, X-ray diffraction, and thermogravimetric-differential thermal analysis. The result indicated that sodium ions entered vermiculite interlayers through the exchange of interlamellar calcium ions. The effects of the heating time on the expansion ratio of Raw-V and Na-V samples were investigated at the temperature range of 400-700 °C. The result indicated that the maximum increment in the expansion ratio could reach up to 26.5% after Na+ modification. The influencing mechanism of Na+ modification on the thermal expansibility of vermiculite was explored via molecular dynamics simulation and the binding energy and dehydration enthalpy change calculation. The simulation and calculation results showed a good agreement with the expansion experiment result. This study provides a novel method for the preparation of high-performance expanded vermiculite.

Study of the interactions of a novel monoclonal antibody, mAb059c, with the hPD-1 receptor.

Programmed cell death 1 (PD-1) monoclonal antibodies have been approved by regulatory agencies for the treatment of various types of cancer, and the mechanism involves the restoration of T cell functions. We report herein the X-ray crystal structure of a fully human monoclonal antibody mAb059c fragment antigen-binding (Fab) in complex with the PD-1 extracellular domain (ECD) at a resolution of 1.70 Å. Structural analysis indicates 1) an epitope, comprising fragments from the C'D, BC and FG loops of PD-1, contributes to mAb059c interaction, 2) an unique conformation of the C'D loop and a different orientation of R86 enabling the capture of PD-1 by the antibody complementarity determining region (CDR) and the formation of one salt-bridge contact - ASP101(HCDR3):ARG86(PD-1), and 3) the contact of FG with light chain (LC) CDR3 is maintained by a second salt-bridge and two backbone hydrogen bonds. Interface analysis reveals that N-glycosylation sites 49, 74 and 116 on PD-1 do not contact mAb059c; while N58 in the BC loop is recognized by mAb059c heavy chain CDR1 and CDR2. Mutation of N58 attenuated mAb059c binding to PD-1. These findings and the novel anti-PD-1 antibody will facilitate better understanding of the mechanisms of the molecular recognition of PD-1 receptor by anti-PD-1 mAb and, thereby, enable the development of new therapeutics with an expanded spectrum of efficacy for unmet medical needs.

Forkhead box N4 (Foxn4) activates Dll4-Notch signaling to suppress photoreceptor cell fates of early retinal progenitors.

The generation of diverse neuronal types and subtypes from multipotent progenitors during development is crucial for assembling functional neural circuits in the adult central nervous system. During mouse retinogenesis, early retinal progenitors give rise to several cell types, including ganglion, amacrine, horizontal, cone, and rod cells. It is unknown at present how each of these fates is selected from the multiple neuronal fates available to the early progenitor. By using a combination of bioinformatic, genetic, and biochemical approaches, we investigated the mechanism by which Foxn4 selects the amacrine and horizontal cell fates from multipotential retinal progenitors. These studies indicate that Foxn4 has an intrinsic activity to suppress the alternative photoreceptor cell fates of early retinal progenitors by selectively activating Dll4-Notch signaling. Gene expression and conditional ablation analyses reveal that Dll4 is directly activated by Foxn4 via phylogenetically conserved enhancers and that Dll4 can partly mediate the Foxn4 function by serving as a major Notch ligand to expand the progenitor pool and limit photoreceptor production. Our data together define a Foxn4-mediated molecular and signaling pathway that underlies the suppression of alternative cell fates of early retinal progenitors.

Inhibition of multiple gene expression and virus replication of HBV by stable RNA interference in 2.2.15 cells.

BACKGROUND/AIMS: Hepatitis B virus (HBV) infection is a world-wide health problem. Recent studies have demonstrated the efficacy of RNA interference (RNAi) against HBV replication at cell culture and animal levels using transient transfection. The present study was to determine whether the stable transfection of short hairpin RNA (shRNA)-producing vector could achieve potent and sustained inhibition of the HBV replication in 2.2.15 cells. METHODS: shRNA-producing vector against HBV and the empty vector were stably transfected into the 2.2.15 cells respectively. A series of experiments were performed in the producing stable lines to determine the changes of viral protein expression and replication. RESULTS: The HBV protein expression and viral replication were suppressed dramatically and stably by the integrated shRNA-producing vectors. Most importantly, this suppression effect persists after 30 passages. CONCLUSIONS: Our data provided the possibility of continuous and stable inhibition of HBV protein expression and replication in patients using RNAi, suggesting a potential clinical application of this novel approach. Furthermore, the established stable transfected cell lines provided a good platform for understanding the mechanism of anti-HBV by RNAi.