Hepatitis B virus （HBV） exclusively infects liver parenchymal cells and forms covalently closed circular DNA （cccDNA） within their nuclei. HBV cccDNA serves as the essential template for viral gene transcription， the sole source of progeny virus production， and the key driver of viral antigen expression， and it is the molecular basis for the persistence of HBV infection. Therefore， elimination and/or functional silencing of cccDNA is the key to eradicate chronic HBV infection. This article discusses the critical scientific issues that need to be solved during elimination of the harm of HBV infection from the perspectives of the synthesis， transcription， and clearance of cccDNA， as well as the impact of nonparenchymal cells on cccDNA， in order to provide a reference for eradicating HBV infection in the future.

Objectives: . Despite the efficacy of surgical treatments, the high rate of recurrence in juvenile nasopharyngeal angiofibroma (JNA) after surgery remains an unresolved problem. The present study comprehensively analyzed the risk factors and characteristics of JNA recurrence, providing clinical guidance for reducing recurrence. Methods: . A total of 123 patients who underwent surgery for JNA between 1997 and 2019 at a single hospital were analyzed retrospectively. Univariate and multivariate analyses were used to assess the clinical risk factors for the recurrence of JNA. The relapse-free survival and annual cumulative recurrence rates were analyzed for subgroups defined according to clinical parameters. Results: . After screening, 78 of the 123 patients were included in the present study. The main risk factors associated with JNA recurrence included the year of diagnosis, tumor size, sphenoid bone invasion, Radkowski stage, surgical approach, and intraoperative bleeding. Importantly, the surgical approach and sphenoid bone invasion were independent prognostic factors affecting recurrence. Patients who underwent endoscopic surgery without sphenoid bone invasion exhibited longer relapse-free survival. In the present study, the overall cumulative recurrence rate of JNA was 38.7%, and recurrence occurred mainly in the first year after the initial surgery. Conclusion . Endoscopic surgery achieved better relapse-free survival in JNA patients, and patients with sphenoid bone invasion should be carefully explored to avoid residual JNA. The recurrence rate of JNA differed among subgroups defined based on clinical parameters and was highest in the first year after surgery. Computed tomography or magnetic resonance imaging, along with close follow-up, should be performed strictly within 1 year after the primary operation.

Objective: To investigate the expression of 11β-hydroxysteroid dehydrogenase (11β-HSD) in polyps of patients with chronic rhinosinusitis with nasal polyps (CRSwNP) and its correlation with glucocorticoid sensitivity. Methods: The prospective study method was applied. Forty-three adult CRSwNP patients from Otorhinolaryngology Hospital, First Affiliated Hospital of Sun Yat-sen University between April 2016 and June 2017 were enrolled in this study. There were 19 males and 24 females with the age of (37.44±7.42) years old. The endoscopic scores by nasal Polyps Grading System before and after one-week prednisone treatment (0.5 mg/(kg·d)) were evaluated. The response of glucocorticoid by the total endoscopic scores was estimated. According to the patient′s reduced nasal polyp endoscopic score, patients were devided into nasal polyps insensitive to glucocorticoids treatment group (insensitive group) and nasal polyp sensitive to glucocorticoids treatment group (sensitive group). The expression of 11β-HSD1, 11β-HSD2 in nasal polyps were measured by Real-time PCR (RT-PCR), Western Blot and immunohistochemisty. According to the clinical data, the Logistic regression models and receiver operation characteristics (ROC) curves were used to explore the predictor for glucocorticoid response in CRSwNP. Results: The expression of 11β-HSD1 and 11β-HSD1/11β-HSD2 was higher in sensitive group than that of insensitive group, while the expression of 11β-HSD2 was lower (rank average was 26.08 vs 16.33, 27.24 vs 14.72, 18.66 vs 26.64, Z value was -2.511, 0.323, -2.059, respectively, all P<0.05). The endoscopic scores in CRSwNP group declined whereas the expression of 11β-HSD1/11β-HSD2 increased (r=0.528, P=0.001), while the cutoff value of the ratio of 11β-HSD1/11β-HSD2 was 2.290 (sensitivity was 79.17%, specificity was 88.89%). Conclusions There is a positive correlation between the response of glucocorticoid and the ratio of 11β-HSD1/11β-HSD2, which could be used as a marker in predicting the level of tissue response to glucocorticoid therapy in CRSwNP.

Objective: To explore the clinical features, diagnostic methods and therapeutic strategy of silent sinus syndrome (SSS). Methods: A retrospective study was made on eight SSS patients treated during 2013-2016 in Longgang ENT Hospital and Otorhinolaryngology Hospital of the First Affiliated Hospital of SUN Yat-sen University. The following clinical data, including demographic data, symptoms, history of trauma and surgery, signs, imaging examination, endoscopic surgery and postoperative outcomes, were analyzed to summarize the diagnosis and treatment experiences. Results: Eight SSS patients showed the following clinical features: the proportions of both sexes and sinus sides were 4 to 4; seven cases (7/8) were adult, with an average of (48.1±11.8)y; seven cases (7/8) had long history of trauma or surgery, with an average of (17.9±10.5)y; seven cases (7/8) cannot recall the exact course of SSS; six cases (6/8) had no nasal symptoms; eight cases (8/8) had unilateral ocular discomforts; eight cases (8/8) had signs of unilateral enophthalmos (2-5 mm), accompanied with hypoglobus; and by CT and MRI scanning, eight cases (8/8) showed the unilateral maxillary sinus outlet obstruction, sinus full opacification, sinus wall bony rarefaction, sinus wall contraction, sinus volume loss, and the ipsilateral orbital floor bowing descent and orbital volume increase. After treated by endoscopic sinus surgery and followed-up for one year, four cases were cured, and the other four improved; no intra- or post-operative complications occurred; and no individual need a two-stage orbital plastic reconstruction. Conclusions SSS often develops in the unilateral maxillary sinus of adult patient with long history of trauma or surgery, but the nasal symptoms and signs are silent. Diagnostic for SSS depends on characteristic ocular sign and sinus CT imaging. Endoscopic sinus surgery helps to improve ocular and nasal signs and promote orbital self-reconstruction.

Objective: To summarize the surgical techniques, benefits and limitations of transnasal endoscopic resection and optic nerve decompression for patients with optic neuropathy caused by fibro-osseous lesions. Methods: Eight patients with optic neuropathy caused by fibro-osseous lesions who accepted endoscopic surgery of either resection of the lesion or decompression of optic nerve in Otorhinolaryngology Hospital, First Affiliated Hospital of Sun Yat-sen University from 2007 to 2016 were retrospectively reviewed and followed until April, 2017. Analyses were performed on the pathology type, disease extent and disease duration, especially on the visual acuity and visual field changes before and after surgery. Results: Eight patients (5 male and 3 female) were included in this study, with a median age of 12 years old (8-19 years old). The median disease duration was 12 months (1-72 months). The visual acuity (VA) of five patients (40 cm/FC, 0.2, 0.1, 0.2, 10 cm/FC, respectively) improved after surgery (0.1, 0.3, 1.2, 0.1, 0.6, respectively), and one patient had no change of VA after the surgery. Two patients (0.02, hand movement, before surgery) became deprived of light perception (VA=0) immediately after surgery. One patient complicated with intra orbital hemorrhage because of anterior artery injury. No complications of cerebral spinal fluid leak, intra-ocular muscle injury, intra-cranial hemorrhage or brain tissue injury occurred. Conclusion For the treatment of optic neuropathy caused by fibro-osseous lesions, transnasal endoscopic surgery might have a good outcome.

Cytoplasmic processing bodies, termed P bodies, are involved in diverse post-transcriptional processes including mRNA decay, nonsense-mediated RNA decay (NMD), RNAi, miRNA-mediated translational repression and storage of translationally silenced mRNAs. Regulation of the formation of P bodies in the context of multicellular organisms is poorly understood. Here we describe a systematic RNAi screen in C. elegans that identified 224 genes with diverse cellular functions whose inactivations result in a dramatic increase in the number of P bodies. 83 of these genes form a complex functional interaction network regulating NMD. We demonstrate that NMD interfaces with many cellular processes including translation, ubiquitin-mediated protein degradation, intracellular trafficking and cytoskeleton structure.We also uncover an extensive link between translation and RNAi, with different steps in protein synthesis appearing to have distinct effects on RNAi efficiency. Moreover, the intracellular vesicular trafficking network plays an important role in the regulation of RNAi. A subset of genes enhancing P body formation also regulate the formation of stress granules in C. elegans. Our study offers insights into the cellular mechanisms that regulate the formation of P bodies and also provides a framework for system-level understanding of NMD and RNAi in the context of the development of multicellular organisms.
Animals ; Animals, Genetically Modified ; Caenorhabditis elegans ; genetics ; Cytoplasmic Structures ; Gene Expression Regulation ; Genes, Helminth ; Genome, Helminth ; genetics ; MicroRNAs ; genetics ; Nonsense Mediated mRNA Decay ; physiology ; RNA Interference ; RNA, Helminth ; genetics ; Reverse Transcriptase Polymerase Chain Reaction

Objective To evaluate the safety of Ad-Mathl administration to inner ear and provide the base data for vestibular dysfunction gene therapy.Methods Ten mature Wistar rats were divided into normal control group(srats) and adenovirus(E1,E3-Deleted and carried mathl and enhanced green fluorescent protein report gene,Ad-Mathl-EGFP)scala vestibuli transfer group(5 rats).Right ears of the Ad-Mathl-EGFP transfering group rats were deliveried 5ul Ad-Mathl-EGFP(physieal tite 2.1 10~(11)v.P./ml)into cochleas through the way of drilling scala vestibuli of cochlear basal turn.As a control,the normal group received nothing to inner ear.In order to estimate functional condition of vestibule and cochlea,the click-evoked potentials on the surface of the cervical dura mater(CDM-CEP),auditory brain stem response(ABR)and swimming time were recorded in all rats at 7 days after treatment,and then histologic and morphologic observation were carried out after animals were sacrificed.Results All animals' morphologic observation showed that inner ear hair cells were normal after transfer.Seven days after transfer,the swimming time was 4.0±0.71 s in normal control group and 5.0±0.71 s in scala vestibuli delivery group.The threshold of CDM-CEP and ABR were 85±3.54 dB SPL and 37±4.47 dB SPL in normal control group,and 89±6.52 dB SPL and 40±3.54 dB SPL in Ad-Mathl-EGFP scala vestibuli delivery group,respectively.There was no significant difference existed between control group and Ad-Mathl-EGFP scala vestibuli delivery group.Conclusion The Ad(E1,E3-Deleted)is safe for vestibular and cochlea hair cells and can be used as an ideal vector of gene transfer.

OBJECTIVE: To observe whether bFGF could cross the blood-labyrinth barrier (BLB) after intra-abdominal injection and to establish an experimental basis for its clinical applications. METHOD: Thirty guinea pigs were divided into three groups. Animals in group 1 were administered o I-bFGF, while animals in group 2 and 3 were administered 125 and saline, respectively, via intra-abdominal injection. The both cochlea, blood, liver, brain, thyroid gland and kidney were collected and weighted. A radioimmunoassay analyzer was employed to measure counts per minute (CPM) of each sample, and autoradiography was performed on both cochlea. RESULT: The CPM value of organ samples in the 125I group was higher than that in other groups, and radioactive grain was observed in cochlear samples of this group. In the 125I-bFGF group, blood demonstrated the highest CPM value, while cochlea and brain demonstrated the lowest CPM value, with no radioactive grain observed in cochlear samples. CONCLUSION bFGF has some difficulties in getting across BLB, so the way of bFGF application in clinics need further study.
Animals ; Autoradiography ; Cochlea ; cytology ; metabolism ; Fibroblast Growth Factor 2 ; administration & dosage ; Guinea Pigs ; Injections, Intraperitoneal ; Iodine Radioisotopes ; administration & dosage

Objective To assess the feasibility of adenoviral vectors mediate cochlear gene transfer by postau-ricular microinjection through the round window membrane in mouse. Methods Twelve 5-week old C57BL/6J mice were selected for the study: 8 were implanted with Ad-EGFP by postauricular microinjection through the round window membrane, and 4 with artificial perilymphatic fluid. On postoperative days 5 and 14, the animals were sac-rificed and the surface preparation of cochleae was observed. Results Two animals died after operation. Bright green fluorescence in the cochleae was observed in Ad- EGFP groups. Gene expression on day 14 after operation was higher than that on day 5. However, the control group was free of fluorescence. Oonclusion The postauricular route of the cochlear gene transfer in mice is simple to operate with little side-effect. The technique of transgenic delivery into the inner ear through RWM by mieroinjection is feasible and effective.

OBJECTIVE To investigate the effect of Hath1 on epithelial ridge (GER) cells from postnatal rat cochlear. METHODS An experimental method was developed which allowed the isolation and culture of GER cells from P1 rat cochleae using a combinatorial approach of enzymatic digestion and mechanical separation. The dissociated GER cells were cultured in DMEM+10% FBS. The GER explants infected with ad-Hath1 for 3 hours were cultured for 10 days and observed under scanning electron microscope. RESULTS The GER cell cultures tended to attach to the substratum and grow in patches which assume a polygonal morphology similar to that of epithelial cells in medium containing 10% serum. Stereociliary bundle-like structures were observed in the boundary of GER cell patches forced to express Hath1. CONCLUSION That GER cells which are likely hair cell progenitors could be cultured in vitro and generate stereociliary bundle-like structure when forced to express Hath1 suggests that the misexpression of Hath1 probably can induce the predifferentiation of pure hair cell progenitors into hair cells in vitro.