Hearing restoration by gene replacement therapy for a multisite-expressed gene in a mouse model of human DFNB111 deafness.

Gene therapy has made significant progress in the treatment of hereditary hearing loss. However, most research has focused on deafness-related genes that are primarily expressed in hair cells with less attention given to multisite-expressed deafness genes. MPZL2, the second leading cause of mild-to-moderate hereditary deafness, is widely expressed in different inner ear cells. We generated a mouse model with a deletion in the Mpzl2 gene, which displayed moderate and slowly progressive hearing loss, mimicking the phenotype of individuals with DFNB111. We developed a gene replacement therapy system mediated by AAV-ie for efficient transduction in various types of cochlear cells. AAV-ie-Mpzl2 administration significantly lowered the auditory brainstem response and distortion product otoacoustic emission thresholds of Mpzl2-/- mice for at least seven months. AAV-ie-Mpzl2 delivery restored the structural integrity in both outer hair cells and Deiters cells. This study suggests the potential of gene therapy for MPZL2-related deafness and provides a proof of concept for gene therapy targeting other deafness-related genes that are expressed in different cell populations in the cochlea.

Synthesis and Evaluation of [18F]AlF-NOTA-c-DVAP: A Novel PET Probe for Imaging GRP78 in Cancer.

GRP78, a member of the HSP70 superfamily, is an endoplasmic reticulum chaperone protein overexpressed in various cancers, making it a promising target for cancer imaging and therapy. Positron emission tomography (PET) imaging offers unique advantages in real time, noninvasive tumor imaging, rendering it a suitable tool for targeting GRP78 in tumor imaging to guide targeted therapy. Several studies have reported successful tumor imaging using PET probes targeting GRP78. However, existing PET probes face challenges such as low tumor uptake, inadequate in vivo distribution, and high abdominal background signal. Therefore, this study introduces a novel peptide PET probe, [18F]AlF-NOTA-c-DVAP, for targeted tumor imaging of GRP78. [18F]AlF-NOTA-c-DVAP was radiolabeled with fluoride-18 using the aluminum-[18F]fluoride ([18F]AlF) method. The study assessed the partition coefficients, stability in vitro, and metabolic stability of [18F]AlF-NOTA-c-DVAP. Micro-PET imaging, pharmacokinetic analysis, and biodistribution studies were carried out in tumor-bearing mice to evaluate the probe's performance. Docking studies and pharmacokinetic analyses of [18F]AlF-NOTA-c-DVAP were also performed. Immunohistochemical and immunofluorescence analyses were conducted to confirm GRP78 expression in tumor tissues. The probe's binding affinity to GRP78 was analyzed by molecular docking simulation. [18F]AlF-NOTA-c-DVAP was radiolabeled in just 25 min with a high yield of 51 ± 16%, a radiochemical purity of 99%, and molar activity within the range of 20-50 GBq/µmol. [18F]AlF-NOTA-c-DVAP demonstrated high stability in vitro and in vivo, with a logD value of -3.41 ± 0.03. Dynamic PET imaging of [18F]AlF-NOTA-c-DVAP in tumors showed rapid uptake and sustained retention, with minimal background uptake. Biodistribution studies revealed rapid blood clearance and excretion through the kidneys following a single-compartment reversible metabolic model. In PET imaging, the T/M ratios for A549 tumors (high GRP78 expression), MDA-MB-231 tumors (medium expression), and HepG2 tumors (low expression) at 60 min postintravenous injection were 10.48 ± 1.39, 6.25 ± 0.47, and 3.15 ± 1.15% ID/g, respectively, indicating a positive correlation with GRP78 expression. This study demonstrates the feasibility of using [18F]AlF-NOTA-c-DVAP as a PET tracer for imaging GRP78 in tumors. The probe shows promising results in terms of stability, specificity, and tumor targeting. Further research may explore the clinical utility and potential therapeutic applications of this PET tracer for cancer diagnosis.

Comparative study of [18F]AlF-PAI-PDL1p and [68Ga]Ga-PAI-PDL1p as novel PD-L1 targeting PET probes for tumor imaging.

PD-L1 is expressed in many tumors but rarely in normal tissues, therefore, it can be a target of PET imaging. In this work, we developed new peptide-based PET probes [18F]AlF-PAI-PDL1p and [68Ga]Ga-PAI-PDL1p with yields of 20-25 % and 40-55 %, respectively. [18F]AlF-PAI-PDL1p and [68Ga]Ga-PAI-PDL1p were synthesized within 30 min with high molar activities. [18F]AlF-PAI-PDL1p and [68Ga]Ga-PAI-PDL1p showed good stability in vivo and in vitro. In vitro cell studies showed [18F]AlF-PAI-PDL1p and [68Ga]Ga-PAI-PDL1p target PD-L1 specifically, with high uptake of 61.52 ± 4.39 and 19.29 ± 2.17 %ID/1 million cells in B16F10 cells at 60 min, respectively. Biodistribution results showed that both [18F]AlF-PAI-PDL1p and [68Ga]Ga-PAI-PDL1p had lower liver accumulation. In vivo PET imaging results showed that [18F]AlF-PAI-PDL1p had a high tumor uptake of 4.23 ± 0.81 %ID/g at 2 h and increased uptake of 6.60 ± 1.01 %ID/g at 12 h. [68Ga]Ga-PAI-PDL1p also showed high tumor uptake of 2.30 ± 0.20 %ID/g at 2 h and slightly increased uptake of 3.80 ± 0.26 %ID/g at 6 h. In conclusion, [18F]AlF-PAI-PDL1p and [68Ga]Ga-PAI-PDL1 seemed to be potential tracers for PET imaging of PD-L1 expression.

Pharmacokinetic analysis of 6-O-[18F]FEE for PET imaging of EGFR mutation.

6-O-[18F]Fluoroethylerlotinib (6-O-[18F]FEE), with a suitable half-life for commercial distribution, may be a good replacement for [11C]erlotinib to identify epidermal growth factor receptor (EGFR) positive tumors with activating mutations to tyrosine kinase inhibitors therapy. In this study, we explored the fully automated synthesis of 6-O-[18F]FEE and investigated its pharmacokinetics in tumor-bearing mice. 6-O-[18F]FEE with high specific activity (28-100 GBq/µmol) and radiochemistry purity (over 99 %) was obtained by two-step reaction and Radio-HPLC separation in PET-MF-2 V-IT-1 automated synthesizer. PET imaging of 6-O-[18F]FEE in HCC827, A431, and U87 tumor-bearing mice with different EGFR expression and mutation was performed. Uptake and blocking of PET imaging indicated that the probe specifically targeted exon 19 deleted EGFR (the quantitative analysis of tumor-to-mouse ratio for HCC827, HCC827 blocking, U87, A431 was 2.58 ± 0.24, 1.20 ± 0.15, 1.18 ± 0.19, and 1.05 ± 0.13 respectively). Dynamic imaging was used to study the pharmacokinetics of the probe in tumor-bearing mice. Logan plot graphical analysis demonstrated late linearity and a high fitting correlation coefficient (0.998), supporting reversible kinetics. According to the Akaike Information Criterion (AIC) rule, the 2-compartment reversible model was more consistent with the metabolic properties of 6-O-[18F]FEE. The automated radiosynthesis and pharmacokinetic analysis will promote clinically transformation of 6-O-[18F]FEE.

Circulating MMP-12 as Potential Biomarker in Evaluating Disease Severity and Efficacy of Sublingual Immunotherapy in Allergic Rhinitis.

Background: Allergic rhinitis (AR) is a highly heterogeneous disease, and allergen-specific immunotherapy (AIT) is an effective treatment. This study aims to evaluate the circulating mas-related G protein-coupled receptor-X2 (MRGPRX2) and matrix metalloproteinase-12 (MMP-12) levels in evaluating disease severity and predicting efficacy of SLIT in AR patients. Methods: We enrolled 110 moderate-severe persist AR patients (AR group) and 40 healthy controls (HC group). Circulating levels of MRGPRX2 and MMP-12 were measured, and their associations with disease severity were evaluated. All AR patients were assigned to receive sublingual immunotherapy (SLIT), and the efficacy was evaluated, and serum samples were collected at 1 year and 3 years after treatment. The correlations between serum MRGPRX2 and MMP-12 and clinical efficacy were assessed. Results: The serum concentrations of MRGPRX2 and MMP-12 were significantly higher in the AR group than the HC group, and the elevated MMP-12 levels were correlated with VAS and TNSS, and serum MRGPRX2 levels were correlated with VAS. Finally, 100 and 80 patients completed 1-year and 3-year follow-up and were classified into effective and ineffective groups. Serum MRGPRX2 and MMP-12 levels were lower in the effective group than the ineffective group. Although serum MRGPRX2 and MMP-12 levels did not significantly change after 1 year SLIT, serum MMP-12 levels were decreased 3 years post-SLIT than baseline and 1 year post-SLIT levels. Receiver operating characteristic (ROC) showed that serum MMP-12 was a potential biomarker for predicting the efficacy of SLIT. Conclusion: Serum MRGPRX2 and MMP-12 appeared to be promising biological indicators in reflecting disease severity in AR patients. Moreover, circulating MMP-12 might serve as a reliable predictor for clinical responsiveness of SLIT.

[The Application Value of True Whole-body PET/CT Scanning Protocol in Patients of Extranodal NK/T Cell Lymphoma].

OBJECTIVE: To assess the staging, restaging, and treatment strategy determination of extranodal NK/T cell lymphoma (ENKT) by PET/CT real body (true whole-body, TWB) imaging, which is superior to PET/CT limitation of the whole body (limited whole-body, LWB, from skull vertex to upper thighs) by adding 'distal lower extremity' images. METHODS: TWB 18F-FDG PET/CT studies performed for staging and follow-up of ENKTL patients between January 2012 and September 2017 were retrospectively reviewed. Patients in staging group received TWB PET/CT evaluation for staging at the first diagnosis. In follow-up group, patients received follow-up evalution with TWB PET/CT and progressive disease (PD) in the LWB range with or without clinical diagnosis or suspicion before follow-up examination, and then divided into four subgroups: staging (+) PD (－), staging (+) PD (+), staging (－) PD (－), staging (－) PD (+). Then the percentage of unexpected ENKTL lesions found at the distal extremity (outside the LWB range) (P1), and the percentage of changes in the staging, restaging/outcome evaluation (P2) in each group were recorded. RESULTS: Among the 225 patients in the staging group, 200 (88.9%) had tumors confined to LWB, while P1 was 11.1% (25 cases) and P2 was 0.4% (1 case). In the follow-up group, the P1 in staging (+) PD (－)( n=85), staging (+) PD (+)( n=4), staging (－) PD (－)( n=43), staging (－) PD (+) goups ( n=15) were 1.2%, 75.0%, 0%, 26.7%, and P2 were 1.2%, 0%, 0%, 13.3%, respectively. In the follow-up group, regardless of whether the TWB PET/CT examination was performed at the initial diagnosis stage, P1 in PD (－) group and PD (+) group was 0.8 vs. 36.8% ( P<0.000 1), and P2 was 0.8% vs. 10.5% ( P<0.000 1). CONCLUSION: It is not recommended that the TWB PET/CT imaging from the top of the head to the bottom of the foot use for the first diagnosis of ENKTL patients. And for follow-up patients with no clinical evidence of tumor progression or with evidence of tumor progression but whose lesions were limited to LWB at the initial diagnosis of TWB PET/CT staging, LWB PET/CT from the top of the head to the middle of the thigh is recommended for routine follow-up. For ENKTL patients, TWB PET/CT was not performed at the initial stage of diagnosis to detect the condition of lower limbs. If the evidence of tumor progression in the LWB range appeared before the follow-up examination, TWB PET/CT was recommended for the follow-up evaluation to evaluate the systemic tumor involvement.

Micro-CT study of the human cochlear aqueduct.

OBJECTIVE: The anatomic structure of the cochlear aqueduct (CA) in human temporal bone specimens was observed using micro-computed tomography (CT). MATERIALS AND METHODS: Micro-CT scanning of 18-µm-thick slices was performed on 30 slides of human temporal bone specimens to observe the CA structure and its relationship with its surroundings. The length, internal and external apertures, and the narrowest width of the CA were measured. The differences in CAs were compared between high jugular bulb (HJB) specimens and normal specimens. RESULTS: A large number of CA images were acquired using Micro-CT scanning, which clearly displayed the basic anatomic structures, stereotactic localizations, and adjacent relationships of the CAs. The whole course of a CA was 12.31 ± 3.60 mm, the diameter of the internal aperture was 465 ± 242 µm, the diameter of the external aperture was 2.88 ± 1.06 mm, the narrowest diameter was 601 ± 335 µm, the diameter of the opening of inferior cochlear vein (ICV) was 151 ± 50 µm, the distance between the internal aperture and ICV was 270 ± 197 µm, and the distance between the inferior margin of the internal acoustic meatus (IAM) and the top most part of the external aperture of the CA was 6.783 ± 2.15 mm. No bony obstruction of the CA or CA enlargement was observed in the specimens. A total of 28 CAs had one accompanying bony canal in the surroundings. The length and travelling of the CA were not affected by the level of the jugular bulb (JB). The variation of the travelling of the ICV was larger than that of the CA. CONCLUSION: Micro-CT adequately displayed the bony CA canal and provided a new method for anatomical studies of the CA and a basis for functional studies.

A novel technique C-conchoplasty in canal wall down tympnomastoidectomy.

BACKGROUND: Numerous methods for meatoplasty and conchoplasty have been introduced, but no clear V/S(the meatal cavity volume to the cross-sectional) was given and many patients have complained about poor cosmesis on follow-up. AIMS: To explore the proper size, and cosmetic shape of the external auditory meatus and auditory canal for canal wall-down tympnomastoidectomy (CWD). MATERIAL AND METHODS: In this observational case series study 36 patients undergone CWD with C-conchoplasty that uses a C-shape skin incision on the concha were reviewed. S and V/S of the preoperative, postoperative and contralateral normal ears were observed. We analyzed the relationship between the epithelialization time and postoperative V/S. Long-term efficacy observation and the shape of the meatus after the operation were observed. RESULTS: C-conchoplasty could effectively enlarge S and reduce V/S. The postoperative V/S were closer to the normal ear than that if we didn't do C-conchoplasty. The greater difference of V/S between the post-operative ears and the contralateral normal ears, the longer the epithelialization time will be. C-conchoplasty produced an excellent cosmetic result. No other complications were noted. CONCLUSIONS AND SIGNIFICANCE: The C-conchoplasty, which is a novel and easy technique in CWD, offers good functional and excellent cosmetic results with minimal risk of complications.

Biomimetic graphene oxide quantum dots nanoparticles targeted photothermal-chemotherapy for gastric cancer.

Direct use of chemotherapy drugs in the treatment of gastric cancer often leads to systemic side effects and unsatisfied therapeutic efficacy due to the lack of tumour-targeting ability. The excellent properties of nanoparticles make them good tools to provide more options for the targeted delivery of chemotherapeutic drugs. Herein, we developed a novel nanomedicine (GOQD-ICG-CS-6@HM nanoparticles, GIC@HM NPs), which employed graphene oxide quantum dots (GOQDs) to co-load photosensitizer indocyanine green (ICG) and chemotherapeutic drug gamabufotalin (CS-6) as the core and wrapped with the hybrid membrane (erythrocyte membrane and gastric cancer cell membrane, HM) on its surface. This nanomedicine possesses the functions of photothermal therapy and chemotherapy, making it a good choice for the treatment of gastric cancer. The results showed that the bionic-coated hybrid membrane not only improves the biocompatibility of the nanomedicine, and prolong its circulating half-life, but also delivers the drug to the tumour site precisely and improves the efficiency of drug utilisation. In vitro and in vivo studies further showed that GIC@HM NPs exhibited combinational effects on tumour therapy while displaying no obvious side effects on normal tissue. To sum up, the newly developed GIC@HM NPs provide a safer, more efficient, and more precise method for gastric cancer treatment.

Mechanisms and otoprotective strategies of programmed cell death on aminoglycoside-induced ototoxicity.

Aminoglycosides are commonly used for the treatment of life-threatening bacterial infections, however, aminoglycosides may cause irreversible hearing loss with a long-term clinical therapy. The mechanism and prevention of the ototoxicity of aminoglycosides are still limited although amounts of studies explored widely. Specifically, advancements in programmed cell death (PCD) provide more new perspectives. This review summarizes the general signal pathways in programmed cell death, including apoptosis, autophagy, and ferroptosis, as well as the mechanisms of aminoglycoside-induced ototoxicity. Additionally, novel interventions, especially gene therapy strategies, are also investigated for the prevention or treatment of aminoglycoside-induced hearing loss with prospective clinical applications.

[Clinical research on the surgical treatment of complex maxillary sinus diseases by pyriform aperture extend prelacimal recess approach].

Objective:The aim of this study is to explore the operative method, indication and clinical effect of enlarging anterior lacrimal Fossa via Piriform foramen for complex lesions of maxillary Sinus. Method:By imaging and pathology, twenty-one patients with tumors and bone cysts were confirmed that the lesion base was located in the anterior wall of maxillary sinus, alveolar recess and zygomatic recessor even more broadly. All patients were treated with pyriform aperture extend prelacimal recess approach surgery, and followed up for average 25 months. Result:All the 21 operations went smoothly, including 11 cases diagnosed with papilloma, 4 cases diagnosed with maxillary sinus carcinoma, 1 case diagnosed with moderate dysplasia, 1 case diagnosed with fibroangioma, 1 case diagnosed with schwannomas, 1 case diagnosed with maxillary sinus osteoma, 1 case diagnosed with maxillary sinus bone cyst and 1 case diagnosed with dentigerous cyst. Pyriform aperture extend prelacimal recess approach can thoroughly deal with lesions located in each part of the maxillary sinus. Intraoperative blood loss（191.6±44.7） mL. After operations, 6 patients had mild facial swelling, and 8 cases experienced facial numbness; There were no facial scar, collapse and deformation, anterior nares narrowing, tears overflow and other complications. None of the patients relapsed. Conclusion:The enlarged anterior lacrimal fossa via piriform aperture can provide enough space for complete resection of the complex lesions of maxillary Sinus, which are located in anterior wall, Alveolar recess, zygomatic recess and various parts of the base, the postoperative reaction is light, the complication is few.

Effect of revascularization on IL-6 and TNF-α in patients with thromboangiitis obliterans.

The aim of the study was to investigate the effects of revascularization in treating patients with thromboangiitis obliterans (TAO), to analyze the prognosis of TAO. The treatment group comprised 32 patients with TAO of lower limbs who were selected between March 2012 and March 2017. Patients in the treatment group were treated with revascularization (vascular bypass surgery, catheter-directed thrombolysis and angioplasty, endovascular angioplasty + stening, thromboectomy and/or endarterectomy) + Western medicine. Another 33 patients with TAO who were treated with Western medicine alone comprised the control group. Treatment outcomes were compared between the groups. Serum levels of interleukin-6 (IL-6), interleukin-8 (IL-8) and tumor necrosis factor-α (TNF-α) were also detected and compared between the groups. Multivariate analysis was performed to identify the factors related to prognosis. Compared with control group, treatment outcomes were significantly better in the treatment group (P<0.05). After treatment, the serum levels of IL-6, IL-8 and TNF-α significantly decreased in both groups, and the decrease in the treatment group was more significant (P<0.01). Multivariate analysis revealed that vascular bypass surgery and preoperative ischemic degree are associated with treatment effect. Our results show that revascularization treatment of TAO is conducive to clinical symptoms and dysfunction of inflammatory cytokines, and the type of surgery and surgical timing significantly affect treatment outcomes.

Immunomodulatory Effect of Flavonoids of Blueberry (Vaccinium corymbosum L.) Leaves via the NF-κB Signal Pathway in LPS-Stimulated RAW 264.7 Cells.

OBJECTIVE: This study aimed to explore the immunoregulatory effect of flavonoids of blueberry (Vaccinium corymbosum L.) leaves (FBL). METHODS: The flavonoids of blueberry leaves were prepared with 70% ethanol and were identified by ultraperformance liquid chromatography/quadrupole-time-of-flight mass spectrometry (UPLC/Q-Tof-MS). The immunoregulatory effect and possible regulatory mechanisms of FBL were investigated in lipopolysaccharide- (LPS-) induced RAW 264.7 cells. RESULTS: According to the results of UPLC/Q-Tof-MS, nine flavonoids of blueberry leaves were identified. FBL showed a significant reduction in the production of TNF-α in LPS-stimulated RAW 264.7 cells. FBL significantly decreased the expression of NF-κB p65 and P-NF-κB p65 in LPS-induced RAW 264.7 cells in a dose-dependent manner. CONCLUSION: Our study showed the immunoregulatory effect of FBL through the suppression of TNF-α via the NF-κB signal pathway.

[Clinical analysis of invasive laryngeal mycosis].

OBJECTIVE: To summarize the clinical datas of thepatients with invasive laryngeal fungal infections in, discuss pathogenesis and treatment methods. METHOD: Eleven cases of invasive laryngeal fmycosis who were collected from September 2006 to February 2010 with electronic laryngoscopy, aspirate smear and culture and tissue biopsy for pathological diagnosis, were restrospectively analyzed. Those patients were received iv fluconazole, treatment of Oxygen Atomization of amphotericin B solution and taking itraconazole orally. The hepatic and renal functions of the patients were monitored in the course of treatment. RESULT: All the cases were diagnosed of invasive laryngeal mycosis. 1 patient showed liver dysfunction in the second week during treatment. And continuing the treatment after using liver protection drugs. All symptoms of the patients were improved and no recurrence happened during the 1-6 years of follow-up. CONCLUSION: Invasive laryngeal fmycosis was correlated with occupation exposure, abusing of antibiotics and low immunity. Laryngeal mycosis was Diagnosised mainly depended on the pathological examination. The positive rates of the secretion smear was low. The effects of iv fluconazole, Oxygen Atomization of amphotericin B 2-4 weeks, and 4 weeks of taking itraconazole orally were safety and reliable.

[Short-term results of audiological change of nasopharyngeal carcinoma patient after radiotherapy and chemotherapy].

OBJECTIVE: To investigate short-stage results of audiological change of nasopharyngeal carcinoma patients after radiotherapy and chemotherapy treatment. METHODS: According to treatment modus of nasopharyngeal carcinoma, 64 cases (128 ears) patients were divided to simple radiotherapy group (45 cases, 90 ears) and radiotherapy with chemotherapy group (combination treatment group, 19 cases, 38 ears). Meanwhile, 25 cases (50 ears) people took as control group, who had no obviously ear and nose disease. About two or three months after radiotherapy and chemotherapy treatment completion, three groups were detected by otoscopy, pure tone test, tympanogram and eustachian tube function, respectively. Then, hearing variation of all patients after radiotherapy and chemotherapy treatment were investigated and compared each other. At the same time, the character and level of audiological change were also analyzed. RESULTS: Eardrum character of nasopharyngeal carcinoma patients appeared change after radiotherapy. Simple radiotherapy and combination treatment groups were found having hearing impairment and eustachian tube functional disturbance. Moreover, most patients of simple radiotherapy group showed conductive deaf (24%, 22/90), and combination treatment group exhibited mingle (24%, 9/38) or sensorineural deafness (29%, 11/38). CONCLUSIONS: Recent hearing of nasopharyngeal carcinoma patients were damaged by radiotherapy and chemotherapy treatment, radiotherapy treatment induced middle ear or eustachian tube function disturbance, chemotherapy treatment had cochleotoxicity, compared with other treatment, combination treatment was more aggravated hearing impairment.