Characteristics of dust mite sublingual immunotherapy-associated adverse events in the early phase.

Background: Few studies reported the characteristics of house dust mite (HDM) sublingual immunotherapy (SLIT) adverse events (AEs) during early phase treatment. The aim of this prospective study was mainly to explore the characteristics of AEs in allergic rhinitis (AR) patients during 6 months of HDM SLIT. Methods: A total of 242 patients with AR were enrolled in this study. Telephone follow-up and administration were conducted in the every week of the first month, the third month, and the sixth month of SLIT treatment. Furthermore, the early efficacy, AEs, and compliance were analyzed in our study. Results: Overall, 70.25% (170/242) of the AR patients completed the study, while 29.75% (72/242) of the AR patients failed to complete the whole 6 months of SLIT treatment process. On the whole, symptoms improved in 87.65% (149/170) of patients including 34.12% (58/170) well-controlled and 53.53% (91/170) partially controlled. The correlation analysis results showed that the treatment effect was negatively correlated with the age (r = -0.1614, P = 0.0355). The AEs mainly occurred in the first month, comprised of local rashes, gastrointestinal reactions, and itching of mouth and tongue. Subgroup analysis in the first month showed the itching of mouth and tongue, gastrointestinal reactions, fatigue, and other AEs in ≥14 years old group (14-65 years old, n = 42) were significant differences when compared with that in the <14 years old group (4-13 years old, n = 128, all P < 0.05). In the study, the main reasons for terminated immunotherapy were drug inaccessibility, loss of follow-up and long course of treatment. Conclusion: Patients with AR who received HDM SLIT revealed an early efficacy after 6 months, with AEs mostly occurred in the first month.

[Preparation and characterization of Ad-ERα-36-Fc-GFP].

ERα-36 is a novel subtype of estrogen receptor α which promotes tumor cell proliferation, invasion and drug resistance, and it serves as a therapeutic target. However, only small-molecule compounds targeting ERα-36 are under development as anticancer drugs at present. Gene therapy approach targeting ERα-36 can be explored using recombinant adenovirus armed with decoy receptor. The recombinant shuttle plasmid pDC316-Ig κ-ERα-36-Fc-GFP was constructed via genetic engineering to express an Ig κ-signaling peptide-leading secretory recombinant fusion protein ERα-36-Fc. The recombinant adenovirus Ad-ERα-36-Fc-GFP was subsequently packaged, characterized and amplified using AdMaxTM adenovirus packaging system. The expression of fusion protein and functional outcome of Ad-ERα-36-Fc-GFP transduction were further analyzed with triple-negative breast cancer MDA-MB-231 cells. Results showed that the recombinant adenovirus Ad-ERα-36-Fc-GFP was successfully generated. The virus effectively infected MDA-MB-231 cells which resulted in expression and secretion of the recombinant fusion protein ERα-36-Fc, leading to significant inhibition of EGFR/ERK signaling pathway. Preparation of the recombinant adenovirus Ad-ERα-36-Fc-GFP provides a basis for further investigation on cancer gene therapy targeting ERα-36.

Treatment of Postinfectious Olfactory Dysfunction Using Corticosteroids.

OBJECTIVE: This study aimed to analyze the efficacy of different doses and forms of corticosteroids in the treatment of postinfectious olfactory dysfunction and to specify the factors related to the curative effects. METHODS: The medical records of patients diagnosed with postinfectious olfactory dysfunction from January 2018 to May 2019 were reviewed. All patients received different doses of oral corticosteroids for 12 days or inhaled corticosteroids for 1 month. RESULTS: A review of the records of 43 patients showed that the mean follow-up time was 6.66 months, and the overall effective rate of treatment was 48.8%. Postinfectious olfactory dysfunction exhibited distinct seasonality, with the highest incidence in March. The curative effect was significantly related to the olfactory discrimination threshold at the first consultation (P = .022), but not significantly associated with age (P = .325), sex (P = .924), duration of disease (P = .526), follow-up time (P = .489), or treatment program (P = .330). CONCLUSIONS: Different doses and forms of corticosteroids showed no significant difference in the therapeutic effect on postinfectious olfactory dysfunction. The olfactory discrimination threshold at the first consultation was the most important factor influencing the prognosis.

Hyperthermal liquid, spray, and smog may be potential risk factors for recurrent laryngeal nerve thermal injury during thyroid surgeries.

PURPOSE: The hyperthermal liquid, spray and smog caused by energy-based surgical devices (EBDs) may be the potential risk factors for recurrent laryngeal nerve (RLN) thermal injury, but not been reported previously. We aim to demonstrated it both in thyroid surgeries and animal experiments. METHODS: 321 thyroid patients and 32 rats were divided into two groups respectively. A type of endoscopic gauze was implemented in the protected group to prevent nerve from thermal risks. Electromyography and laryngoscope were recorded to determine nerve's conductive ability. Microstructural morphological damage to nerves and muscles were evaluated by light and electron microscopy in the animal experiments. RESULTS: After thyroid surgeries, the incidence of transient vocal cord (VC) palsy was decreased in the protected group (1.4%) compared with the conventional group (4.7%) (P = 0.037). In animal experiments, the average nerve conductive velocity was significantly decreased in the conventional group (5.8 ± 2.1 vs. 25.5 ± 4.7 m/s, P < 0.01). Typical acute morphological thermal changes of edema, coagulation and homogenization in nerve tissues were recorded by light microscopy in the conventional group. Four weeks later, the average weight of gastrocnemius muscles in the injured limb was significantly decreased (1.3 ± 0.2 vs. 2.2 ± 0.2 g, P < 0.01), while no significant difference was noted in the protected group (2.0 ± 0.3 vs. 2.3 ± 0.4 g, P = 0.61). CONCLUSIONS: Hyperthermal liquid, spray or smog may represent risk factors contributing to RLN thermal injury caused by EBDs. Damage to the nerve's conductive ability and morphological changes in nerves and muscles likely represent typical responses to thermal injury. Implementation of endoscopic gauze may be a promising method to protect the RLN from thermal damage especially in endoscopic thyroid surgeries.

Effective inhibition of cancer cells by recombinant adenovirus expressing EGFR-targeting artificial microRNA and reversed-caspase-3.

The EGFR-targeting cancer therapies are commonly facing drug resistance, mostly due to mutations. Gene therapy with artificial microRNA targeting EGFR conserved sequence may avoid such problem. In this study, we constructed a recombinant adenovirus expressing EGFR-targeting artificial microRNA and active revCASP3 (Ad-EC), under the control of tumor-specific SLPI promoter, and evaluated its inhibitory effect on HEP-2 cancer cells both in vitro and in vivo. MTT assay showed that cell growth inhibition rate at 72h was 44.0% in Ad-EC group at MOI 50, while the rate was 7.7% in the control virus Ad-GFP group and 3.6% in Cetuximab (500 µg/ml) group respectively. Flow cytometry analysis revealed the late apoptotic cells rate was 36.1% in Ad-EC group, significantly higher than 6.5% of Ad-GFP group (p < 0.001). When Ad-EC (MOI 50) was combined with CDDP (0.25 µg/ml), late apoptotic cells rate increased to 61.2%, significantly higher than each monotherapy group (P < 0.001). The real-time xCELLigence system recorded an effective cell growth inhibition in Ad-EC and CDDP groups, and more enhanced effect in Ad-EC plus CDDP group. Western blot revealed that Ad-EC could inhibit the activation of AKT pathway and ERK1/2 pathway, while Cetuximab had the AKT pathway over-activated. In vivo experiments with HEP-2 xenograft in nude mice confirmed the tumor inhibition in Ad-EC, CDDP and Ad-EC plus CDDP groups compared with PBS group (P < 0.01). Collectively, these data support the effective inhibition of cancer cells by this novel gene therapy strategy.

Detection of Mitochondrial Mutations Through Isothermal Nucleic Acid Amplification Coupled With Clustered Regularly Interspaced Short Palindromic Repeat-Associated Endonuclease Cas13a.

The clustered regularly interspaced short palindromic repeat (CRISPR)-associated endonuclease Cas13a can specifically bind and cleave RNA. After nucleic acid pre-amplification, bacterial Cas13a has been used to detect genetic mutations. In our study, using a transcription-mediated amplification together with Cas13a, we can isothermally amplify and detect mitochondrial point mutations under non-denaturing conditions from human genomic DNA. Unlike previous reports, we prepared CRISPR DNA with T7 promoter sequences and generated CRISPR RNA via transcription-mediated amplification instead of synthesizing and adding CRISPR RNA in a separate step. As a proof-of-concept, we showed that both m.1494C > T and m.1555A > G mutations were detected within 90 min. In addition, we explored various designs of CRISPR DNA to improve assay specificity, including the location and number of nucleotide mismatches, length of protospacer sequence, and different buffering conditions. We also confirmed the possibility of a "one-step single-tube" reaction for mutation detection. This assay can robustly distinguish circular DNA templates that differ by a single nucleotide. It has the potential to be adapted for automated applications, such as the screening of mitochondrial diseases.

IL-35 inhibits CD8+ T cells activity by suppressing expression of costimulatory molecule CD28 and Th1 cytokine production.

BACKGROUND: Interleukin-35 (IL-35), a novel immune-suppressing cytokine, can promote tumor angiogenesis and inhibits anti-tumor cytotoxic lymphocyte response. Here, we aimed to investigate the potential mechanism of the effects of IL-35 on anti-tumor cytotoxic lymphocyte. METHODS: Dendritic cells (DCs) were used to induce anti-tumor cytotoxic lymphocyte. Flow cytometry, carboxyfluorescein succinimidyl ester staining, ELISA assay and western blotting were used to analyze the effect of IL-35 on anti-tumor cytotoxic lymphocyte. RESULTS: We observed that IL-35 inhibited the expression of costimulatory molecule CD28 on CD8+ T cell surface and Th1 cytokine production. However, IL-35 did not inhibit anti-tumor cytotoxic lymphocyte proliferation nor enhance the expression of apoptosis-related proteins of anti-tumor cytotoxic lymphocyte. Moreover, IL-35 did not repress the expression of Fas ligand (FasL) on cytotoxic lymphocyte surface. CONCLUSIONS: Our findings revealed that IL-35 can inhibit CD8+ T cells activity by suppressing the expression of costimulatory molecule CD28 and Th1 cytokine production.

IL-35 expression is increased in laryngeal squamous cell carcinoma and in the peripheral blood of patients.

Interleukin-35 (IL-35) has been proposed as a novel immune-suppressing cytokine. However, the function of IL-35 in malignant diseases is yet to be elucidated. The present study investigated IL-35 expression levels in laryngeal squamous cell carcinoma (LSCC) tissues and the peripheral blood of patients to explore the potential involvement of IL-35 in LSCC progression. In the present study, IL-35 expression levels in tissues and peripheral blood were analyzed by reverse transcription-quantitative polymerase chain reaction and an enzyme-linked immunosorbent assay. The association between IL-35 expression levels and clinical characteristics was also evaluated. The present results demonstrated that IL-35 expression in tumor tissues was significantly higher than in adjacent normal tissues, and a significant association between IL-35 expression levels in tissues and the tumor site was detected. Furthermore, the expression of IL-35 in the peripheral blood of patients was significantly decreased subsequent to tumor resection. No correlation between peripheral blood IL-35 expression and clinical characteristics was detected. In conclusion, the present study demonstrated that IL-35 is highly expressed in LSCC tissues and in the peripheral blood of patients with LSCC. There was a notable, significant reduction of peripheral blood IL-35 expression following surgical resection of tumors. These results may be useful for diagnostic or therapeutic purposes in patients with LSCC.

Biodegradable nasal packings for endoscopic sinonasal surgery: a systematic review and meta-analysis.

OBJECTIVES: To assess biodegradable nasal packing effectiveness for improving postoperative symptoms and mucosal healing after endoscopic sinonasal surgery as compared with conventional/non-packing groups. METHODS: Relevant articles were searched on PubMed, EMBASE, and the Cochrane Central Register of Controlled Trials. We included randomized controlled trials (RCTs) that compared biodegradable packings with conventional packings or no packing, reporting postoperative symptoms and/or mucosal healing outcomes. RESULTS: This review included 19 studies, of which 11 compared biodegradable packings with conventional packings. Meta-analysis found that biodegradable packings significantly improved postoperative symptoms: bleeding at removal, pain at removal, pain in situ, and nasal blockage. Mucosal healing outcomes were inconsistent within studies, with no data could be pooled. Eight studies compared biodegradable packings with non-packing group. Postoperative symptom data in this comparison could not be pooled: A protective or equal effect on postoperative bleeding was reported in different studies; no difference was reported in pain status and nasal blockage. As for mucosal healing, meta-analysis showed that two arms of comparison had similar effect on synechiae, edema, infection and granulation at each time point. CONCLUSION: The limiting evidence suggests that biodegradable nasal packings are statistically better than conventional packings in postoperative symptoms, and probably comparable to non-packing group, as in this comparison we could not carry out meta-analysis. No beneficial or detrimental effect on postoperative mucosal healing could be determined based on existing evidence.

[Longterm impact on swallowing quality-of-life after partial laryngectomy].

OBJECTIVE: To discuss the longterm quality-of-life related to swallowing function after different partial laryngectomy. METHODS: The worldwide known swallow quality-of-life questionnaire (SWAL-QOL, Hongkong, Chinese edition); was used in this research to evaluate the swallowing QOL on 96 postoperative patients who underwent different kinds of partial laryngectomy more than one year before. The patients were divided into 4 groups: supracricoid partial laryngectomy-cricohyoidopexy (SCPL-CHP), supracricoid partial laryngectomy-cricohyoidoepiglottopexy (SCPL-CHEP), horizontal supraglottic partial laryngectomy (horizontal PL), and vertical partial laryngectomy (vertical PL). RESULTS: A one-way MANOVA revealed a significant multivariate (the 11 scales of SWAL-QOL) main effect for groups (P < 0.01), and significant univariate main effects were obtained for groups in 9 scales out of 11 (P < 0.01). In all the 9 scales vertical PL group acquired near full scores except the communication scale, and was significantly higher than the other 3 groups in many scales (P < 0.05). CHP group acquired the worst scores of the 4 groups, showing significant differences in most of the 9 scales (P < 0.05). No significant difference was found between Horizontal PL and CHEP except in communication (P > 0.05). Patients with deglutition disorders (choke/cough) had a lower score in the social function scale. CONCLUSIONS: Swallowing quality-of-life of postoperative patients was deeply influenced even when more than one year had passed after surgery. Some of them felt deeply burdened by deglutition disorder. Patients after CHP proved to have a worst quality of life than the others, while vertical PL the best. The QOL between Horizontal PL and CHEP was shown to be almost the same. The influence over QOL from longterm dysphagia was multi-dimensional, containing the degeneration of social function.