Incidence and prognosis of olfactory and gustatory dysfunctions related to SARS-CoV-2 Omicron strain infection in China: A national multicenter survey of 35,566 individuals.

Objective: This cross-sectional study aimed to determine the epidemiology of olfactory and gustatory dysfunctions related to COVID-19 in China. Methods: This study was conducted by 45 tertiary Grade-A hospitals in China. Online and offline questionnaire data were obtained from patients infected with COVID-19 between December 28, 2022, and February 21, 2023. The collected information included basic demographics, medical history, smoking and drinking history, vaccination history, changes in olfactory and gustatory functions before and after infection, and other postinfection symptoms, as well as the duration and improvement status of olfactory and gustatory disorders. Results: Complete questionnaires were obtained from 35,566 subjects. The overall incidence of olfactory and taste dysfunction was 67.75%. Being female or being a cigarette smoker increased the likelihood of developing olfactory and taste dysfunction. Having received four doses of the vaccine or having good oral health or being a alcohol drinker decreased the risk of such dysfunction. Before infection, the average olfactory and taste VAS scores were 8.41 and 8.51, respectively; after infection, they decreased to 3.69 and 4.29 and recovered to 5.83 and 6.55 by the time of the survey. The median duration of dysosmia and dysgeusia was 15 and 12 days, respectively, with 0.5% of patients having symptoms lasting for more than 28 days. The overall self-reported improvement rate was 59.16%. Recovery was higher in males, never smokers, those who received two or three vaccine doses, and those that had never experienced dental health issues, or chronic accompanying symptoms. Conclusions: The incidence of dysosmia and dysgeusia following infection with the SARS-CoV-2 virus is high in China. Incidence and prognosis are influenced by several factors, including sex, SARS-CoV-2 vaccination, history of head-facial trauma, nasal and oral health status, smoking and drinking history, and the persistence of accompanying symptoms.

The value of gadolinium-enhanced MRI in predicting the development of sudden hearing loss into Ménière's disease.

OBJECTIVE: To compare the clinical features of sudden hearing loss (SHL) in patients with and without endolymphatic hydrops (EH), and to investigate the association between SHL with EH and Ménière's disease (MD). METHODS: The clinical data of 63 SHL patients with first symptoms were evaluated retrospectively. Patients were separated into two groups based on the results of gadolinium-enhanced magnetic resonance imaging: EH and non-EH groups. Independent sample t-test and U-test were used to compare groups for continuous variables, and the chi-squared test, corrected chi-squared test and Bonferroni correction test were used to compare groups for binary and ordinal variables. The binary logistic regression model was utilised for univariate and multivariate analysis of follow-up patient prognosis. RESULTS: The EH and non-EH groups contained 32 and 31 patients, respectively. The EH group had a higher prevalence of low-tone descending hearing loss. Fifty-one patients were followed for more than 2 years. In the EH group, 11 and 15 patients were diagnosed with sudden sensorineural hearing loss (SSNHL) and MD, respectively, while in the non-EH group, 24 patients were diagnosed with SSNHL and only one with MD. EH, low-tone descending hearing loss and vertigo were risk factors for the diagnosis of MD in a subgroup univariate regression analysis of patients experiencing SHL. EH was found to be a risk factor for the progression of SHL into MD in a multifactor regression analysis. CONCLUSIONS: Patients with SHL who have EH are more likely to present with low-tone descending hearing loss. EH is a risk factor for the subsequent development of MD.

The Hyperproliferation Mechanism of Cholesteatoma Based on Proteomics: SNCA Promotes Autophagy-Mediated Cell Proliferation Through the PI3K/AKT/CyclinD1 Signaling Pathway.

Cholesteatoma is a chronic inflammatory ear disease with abnormal keratinized epithelium proliferation and tissue damage. However, the mechanism of keratinized epithelium hyperproliferation in cholesteatoma remains unknown. Hence, our study sought to shed light on mechanisms affecting the pathology and development of cholesteatoma, which could help develop adjunctive treatments. To investigate molecular changes in cholesteatoma pathogenesis, we analyzed clinical cholesteatoma specimens and paired ear canal skin with mass spectrometry-based proteomics and bioinformatics. From our screen, alpha-synuclein (SNCA) was overexpressed in middle ear cholesteatoma and might be a key hub protein associated with inflammation, proliferation, and autophagy in cholesteatoma. SNCA was more sensitive to lipopolysaccharide-induced inflammation, and autophagy marker increase was accompanied by autophagy activation in middle ear cholesteatoma tissues. Overexpression of SNCA activated autophagy and promoted cell proliferation and migration, especially under lipopolysaccharide inflammatory stimulation. Moreover, inhibiting autophagy impaired SNCA-mediated keratinocyte proliferation and corresponded with inhibition of the PI3K/AKT/CyclinD1 pathways. Also, 740Y-P, a PI3K activator reversed the suppression of autophagy and PI3K signaling by siATG5 in SNCA-overexpressing cells, which restored proliferative activity. Besides, knockdown of SNCA in RHEK-1 and HaCaT cells or knockdown of PI3K in RHEK-1 and HaCaT cells overexpressing SNCA both resulted in attenuated cell proliferation. Our studies indicated that SNCA overexpression in cholesteatoma might maintain the proliferative ability of cholesteatoma keratinocytes by promoting autophagy under inflammatory conditions. This suggests that dual inhibition of SNCA and autophagy may be a promising new target for treating cholesteatoma.

Peripheral Multiple Cytokine Profiles Identified CD39 as a Novel Biomarker for Diagnosis and Reflecting Disease Severity in Allergic Rhinitis Patients.

Background: Allergic rhinitis (AR) is a common clinical problem, and immune cells and cytokines were proven to be pivotal in its pathogenesis. Our aim is to measure the peripheral concentrations of multiple cytokines in AR patients and identify novel biomarkers for diagnosis and disease severity. Methods: Peripheral blood samples were collected from 50 AR patients, including 25 mild AR (MAR) patients and 25 moderate-severe AR patients (MSAR), and 22 healthy controls (HCs), and multiple cytokine profiling was outlined by Luminex assay. Cytokine levels were compared among the three groups, and their correlations with disease severity were evaluated. The candidate cytokines were further verified by enzyme-linked immunosorbent assay (ELISA) in a validation cohort. Results: Multiple cytokine profiling revealed that CD39 and interferon (IFN)-γ levels were reduced, and interleukin (IL)-13, IL-5, IL-33, and thymic stromal lymphopoietin (TSLP) levels were elevated in the AR group than the HC group (P < 0.05). Receiver operating characteristic (ROC) curves presented that serum CD39 and IL-33 exhibited strong diagnostic abilities, and serum CD39 and IL-10 presented capacities in distinguishing disease severity (AUC > 0.8, P < 0.05). Moreover, CD39 concentrations were decreased, and IL-10, IL-5, and TSLP concentrations were enhanced in the MSAR group more than in the MAR group. Correlation analysis results showed that serum CD39, IL-5, and TSLP levels were associated with total nasal symptom score (TNSS) and visual analogue score (VAS) (P < 0.05). Further data in the validation cohort suggested that serum CD39 levels were reduced, and IL-5 and TSLP levels were increased in AR patients, especially in MSAR patients (P < 0.05). ROC results revealed potential values of serum CD39 in diagnosis and disease severity evaluation in AR patients (P < 0.05). Conclusion: This study highlighted that peripheral multiple cytokine profiles were significantly varied in AR patients and associated with disease severity. The results in discover-validation cohorts implied that serum CD39 might serve as a novel biomarker for diagnosing AR and reflecting its disease severity.

The value of 3D quantitative scoring of endolymphatic hydrops in the diagnosis and differential diagnosis of Ménière's disease.

Objective: We aimed to explore the application of three-dimensional (3D) quantitative scoring of endolymphatic hydrops (EH) based on gadolinium-contrast MRI in both the diagnosis and differential diagnosis of Ménière's disease (MD). Methods: Local threshold method based on signal intensities was used to quantitatively measure volumes, and the degree of EH was scored by 3D quantitative scoring. The receiver-operating characteristic (ROC) curve was used to compare the diagnostic efficacy of this method against conventional evaluation methods. The scores of MD, sudden sensorineural hearing loss (SSNHL), delayed endolymphatic hydrops (DEH), and vestibular migraine (VM) group were compared and correlated with clinical examination results, including pure tone audiometry (PTA), electrocochleogram, and caloric test. Results: The sensitivity and specificity of 3D quantitative scoring were 94.1% and 92.2%, while conventional evaluation methods were 84.3% and 88.2%, respectively. Cochlear score (CR), vestibular score (VR), semicircular canal score (SR), and total scores (TR) in MD group were significantly higher than those in SSNHL group. The rate of diagnosis of MD was significantly higher in the MD group (92.2%) than VM group (27.3%). In MD, PTA was significantly correlated with CR and SR, the ratio of summating potential to action potential (-SP/AP) of electrocochleogram was significantly correlated with CR, VR, and SR, also canal paresis (CP) value of caloric test was significantly correlated with SR. Conclusion: 3D quantitative scoring demonstrated better diagnostic efficacy than conventional evaluation methods for Ménière's disease, and it may be an effective clinical tool to distinguish MD from SSNHL and VM. The clinical practicality of inner ear-enhanced MRI was further confirmed. Level of Evidence: IV.

Magnetic resonance imaging of endolymphatic hydrops in Ménière's disease: A comparison of the diagnostic value of multiple scoring methods.

Objectives: To compare three methods of scoring endolymphatic hydrops in patients with Ménière's disease in order to assess the correlation between endolymphatic hydrops and auditory characteristics. Methods: A retrospective study of 97 patients with unilateral definite Ménière's disease (DMD) who underwent contrast-enhanced three-dimensional fluid attenuated inversion recovery (3D FLAIR) MRI. Each patient was scored by the Inner Ear Structural Assignment Method (IESAM), the Saccule to utricle area ratio (SURI), and the Four Stage Vestibular Hydrops Grading (FSVH), according to their corresponding axial images. Cohen's Kappa and intra-class correlation coefficient were used for consistency testing, combined with binary logistic regression analysis, to compare the sensitivity and specificity of the three methods. The degree of hydrops in different stages of MD was compared. The correlation between endolymphatic hydrops in the inner ear sub-units and hearing thresholds was further analyzed. Results: The intra- and inter-reader reliability for the scoring of endolymphatic hydrops were excellent. The IESAM had a high diagnostic value for identifying definite Ménière's disease (sensitivity: 86.6%, specificity: 97.9%). The hearing thresholds were correlated with the degree of endolymphatic hydrops. Stages 3 and 4 were more significant for the severity of hydrops than stage 1. Within the subgroups of the Ménière's disease patients, compared with the non-hydrops group and the pure vestibular hydrops (V group), the cochlear combined vestibular hydrops group (CV group) had significantly higher auditory thresholds. The amplitude ratio of electrocochleogram was significantly higher in the affected ear than in the healthy ear. Conclusion: The IESAM is a more sensitive and specific diagnostic scoring method for the diagnosis of DMD. Diagnostic imaging may improve the detection of inner ear hydrops which is correlated with severity of hearing loss. A comprehensive evaluation of the inner ear sub-unit structures maybe necessary.

The biofilm characteristics and management of skin flap infection following cochlear implantation.

Objective: This study aims to assess the frequency, bacteriology, biofilm characteristics and management of skin flap infection (SFI) following cochlear implantation (CI). Methods: The study enrolled 1,251 patients receiving CI in the First Affiliated Hospital of Fujian Medical University between August 2001 and March 2021. Scanning electron microscopy (SEM) was utilised to characterise the aetiology of infection. A proposed classification system was applied to optimise treatments for post-operative skin flap infection. Results: After CI, SFI was reported in 16 patients (1.28%) and occurred more frequently in patients under 6 years of age. Of all SFI cases Staphylococcus aureus was the most common pathogen for flap infection, with 8 cases (50%) and bacterial biofilm was evident within the jelly-like substance on the surface of implanted devices in SFI patients. A two-stage classification was proposed to optimise the treatment schemes. Conservative therapy was recommended for stage I cases and surgical treatment for stage II patients. Conclusions: Paediatric patients are more susceptible to SFI after CI, which may be attributed to the formation of bacterial biofilm. The proposed classification can facilitate the management of SFI.

Comparative endoscopic techniques of medial rectus muscle retraction for approaching intraconal tumors: Our experience with five cases.

Objective: To examine the role of transnasal endoscopic approaches in the management of intraconal tumors and demonstrate the use of an eyelid speculum in comparison with different techniques of medial rectus muscle (MRM) retraction. Methods: Retrospective data of five patients with intraconal tumors operated on and followed up by the senior authors between December 2019 and April 2022 was collected. Presenting symptoms, technical details, imaging and histology findings, outcomes, and complications were evaluated. Results: Four primary and one recurrent tumor were identified. The mean patient age was 50 (range, 29-64) years. One tumor was located lateral to the optic nerve, one central and three medial. A complete surgical resection was obtained in four primary cases and a partial resection was achieved in the recurrent case. The MRM was retracted using three different techniques: (1) an infant eyelid speculum creating an operative window between the medial and inferior rectus muscle, (2) external MRM disinsertion transconjunctivally, (3) a four-handed technique performed transseptally by two surgeons. Transient postoperative ophthalmoplegia was recorded in four cases and transient ptosis in one. Three patients completely recovered in 2-3 months while one undergoing MRM disinsertion ended up in restricted strabismus at 15-month follow-up. No other long-term complications have been noted in all five patients with a mean follow-up of 22 (range, 14-32) months. No patients with primary tumors have required additional surgery for tumor recurrence. Conclusion: The indication of endoscopic intraconal surgery may expand to lesions lateral to the optic nerve when the nerve is not in its natural position. The well-known advantages of the endoscopic techniques, namely the lack of external scars, better visualization, less bleeding, and fewer complications, were confirmed. An eye speculum provides a better surgical corridor and eases the pressure exerted on the MRM, which has a promising application prospect.

Ultrasensitive quantitation of circulating miR-195-5p with triple strand displacement amplification cascade.

Circulating miR-195-5p has been proposed as a promising peripheral biomarker for the diagnosis, prognosis and severity assessment of various diseases. However, the demand for its sensitive and convenient quantification has not been met yet. Herein, we proposed a one-pot isothermal approach, in which the target signal acquisition, amplification and conversion (fluorescence read-out) system was integrated by a triple strand displacement amplification (SDA) cascade. Using this triple SDA strategy, miR-195-5p can be at least detected at 1 aM, and the linear dynamic range (from 100 aM to 1 pM) is wide enough to meet the detection needs of clinical miRNA level. A proof-of-principle study, using this novel methodology to directly analyze the spiking serum samples with different levels of miR-195-5p, demonstrated the potential of circulating miR-195-5p detection for clinical point-of-care assay. This one-pot isothermal triple SDA approach, we believe, will be a simple and feasible tool for ultrasensitive quantification of circulating miR-195-5p, and may promote the wide application of this potential biomarker in non-invasive clinical diagnosis.

Positive Feedback Regulation of Poly(ADP-ribose) Polymerase 1 and the DNA-PK Catalytic Subunit Affects the Sensitivity of Nasopharyngeal Carcinoma to Etoposide.

Etoposide (VP-16) is used for the treatment of various cancers, including nasopharyngeal carcinoma (NPC); however, cancers develop resistance to this agent by promoting DNA repair. The DNA-PK (DNA-PKcs) catalytic subunit and poly(ADP-ribose) polymerase 1 (PARP1) mediate acquired resistance and poor survival in NPC cells exposed to DNA damaging agents. DNA repair can alter the sensitivity of NPC cells to DNA damaging agents, and these two enzymes function concomitantly in response to DNA damage in vivo. Therefore, we explored the relationship between DNA-PKcs and PARP1, which may affect NPC cell survival by regulating DNA repair after VP-16 treatment. We performed quantitative real-time polymerase chain reaction, western blotting, and enzyme-linked immunoassays and found that DNA-PKcs knockdown downregulated the PARP1 and PAR expression. Conversely, PARP1 knockdown reduced DNA-PKcs activity, indicating the mutual regulation between DNA-PKcs and PARP1 in VP-16-induced DNA repair. Moreover, a combination treatment with olaparib (a PARP1 inhibitor) and NU7441 (a DNA-PKcs inhibitor) sensitized NPC cells to VP-16 in vitro and in vivo, suggesting that the combined treatment of olaparib, NU7441, and a DNA-damaging agent may be a successful treatment regimen in patients with NPC.

C0818, a novel curcumin derivative, induces ROS-dependent cytotoxicity in human hepatocellular carcinoma cells in vitro via disruption of Hsp90 function.

Heat shock protein 90 (Hsp90) is the most common molecular chaperone that controls the maturation of many oncoproteins critical in tumor development. Hsp90 has been considered as a promising target for cancer treatment, but the clinical significance of Hsp90 and the mechanisms of Hsp90 regulating the tumor-promoting effects in hepatocellular carcinoma (HCC) remain obscure. Previous studies have shown that curcumin, a polyphenol derived from the plant turmeric (Curcuma longa), inhibits tumor growth, which may provide an effective alternative therapy for HCC. Compared to curcumin, a novel derivative of curcumin, 3,5-(E)-Bis(3-methoxy-4-hydroxybenzal)-4-piperidinone hydrochloride (C0818) that is more potent in Hsp90 inhibition and antitumor activity. In this study, we investigated the effect of C0818 on HCC cells in vitro and its relation to Hsp90 inhibition. We showed that C0818 concentration-dependently inhibited the proliferation, the colony formation and induced apoptosis in HepG2 and Sk-Hep-1 cells. C0818 concentration-dependently inhibited DNA synthesis and induced G2/M phase arrest in HepG2 and Sk-Hep-1 cells. We further demonstrated that C0818 induced ROS- and caspase-dependent apoptosis in HCC cells through the mitochondrial-mediated pathway. C0818 induced the degradation of Hsp90 client proteins as RAS, C-Raf, P-C-Raf, Erk, P-ERK, MEK, P-MEK, Akt and P-Akt, which led to subsequent inhibition of the RAS/RAF/MEK/ERK and PI3K/AKT pathways. We revealed that C0818 could inhibit the binding of Hsp90 with its clients without affecting their transcription, which subsequently induced the degradation of Hsp90 clients by the proteasome rather than the lysosome. These results are of potential importance for elucidating a novel Hsp90 inhibitor targeting HCC.

Possible mechanism of CHI3L1 promoting tonsil lymphocytes proliferation in children with obstructive sleep apnea syndrome.

BACKGROUND: The typical characteristic of pediatric obstructive sleep apnea syndrome (OSAS) is systemic inflammation and adenotonsillar hypertrophy (ATH), but the inflammatory markers and mechanism of adenotonsillar proliferation are unclear. METHODS: IHC, qPCR, and western blotting were used to identify the expression of CHI3L1 in the tonsils of children with OSAS. The primary tonsil lymphocytes (PTLCs) from children with OSAS were cultured and recombinant human CHI3L1 protein was added to culture media. After the stimulation with CHI3L1 protein of different concentrations and time points, lymphocyte proliferation was assessed by CCK-8 kits and flow cytometry. The activation of ERK1/2 and the effects on the proliferation of PTLCs were observed by western blotting. RESULTS: The expression of CHI3L1 was higher in the OSAS group than in the PS group. CHI3L1 (100 ng/mmol for 24 h) resulted in a significant increase in the proliferation rate. The ERK1/2 activator (PMA) promoted the proliferation of PTLCs and inhibitor AG126 significantly inhibited proliferation. CONCLUSIONS: CHI3L1 can promote the proliferation of tonsil lymphocytes via ERK1/2 pathways. This result indicates that CHI3L1 may play an important role in the pathogenesis of OSAS in children. Inhibition of CHI3L1 or ERK1/2 may be potential therapeutic targets for CHI3L1-induced proliferation in childhood OSAS. IMPACT: CHI3L1 may be an inflammatory marker in childhood OSAS. CHI3L1 can promote the proliferation of PTLCs in a concentration and time-dependent condition. CHI3L1 can promote the proliferation of tonsil lymphocytes via ERK1/2 pathways.

A nationwide survey of otolaryngologists' compliance with Chinese guidelines for diagnosis and treatment of allergic rhinitis.

BACKGROUND: A higher compliance with clinical guidelines helps improve treatment outcomes. But the clinical practice of otolaryngologists is not always consistent with guidelines. OBJECTIVE: To describe otolaryngologists' compliance with guidelines about allergic rhinitis (AR) management and identify factors responsible for the discordance between clinical practice and guideline recommendations in China. METHODS: A cross-sectional nationwide survey was designed and conducted via an online platform. Recruitment was done by emailing otolaryngologists registered in the Chinese Society of Otorhinolaryngology-Head and Neck Surgery or by inviting otolaryngologists to scan a Quick Respond (QR) code that linked to the questionnaire at various academic meetings. RESULTS: A total of 2142 otolaryngologists were eligible and completed the survey. Of them, 64.7% had over 10 years work experience and 97.4% had a bachelor's degree or higher. About 18.3% of the participants strictly copied the guideline in clinical practice, while 73.7% used the guideline that had been adjusted according to their clinical experience. Otolaryngologists were most concerned about the efficacy, safety, and minimum age of AR medications, and least concerned about patient preferences. Regarding the use of intranasal steroids (INS), leukotriene receptor antagonists (LTRA), and H1-antihistamines, 86.8%, 55.7% and 51.2% of otolaryngologists complied with the guideline recommendations, respectively. Educational background was a factor affecting the compliance with guidelines and acceptance of INS. CONCLUSION: A vast majority of Chinese otolaryngologists complied with the current Chinese AR guidelines. A difference still existed between the otolaryngologists' real-world and guideline-recommended management. The otolaryngologists should pay more attention to patient preferences. A higher education could improve otolaryngologists' adherence to the guidelines.

Tuberculous Otitis Media Complicated by Meningitis-Induced Bilateral Sensorineural Hearing Loss: A Case Report.

Tuberculosis of the middle ear is a rare but treatable disease; however, delays in diagnosis and treatment usually lead to complications. Diagnosis is made difficult by most physicians being unfamiliar with the typical presenting features and special cultural and pathologic studies being required for diagnosis. A case report and literature review are presented, illustrating typical clinical, epidemiologic, and laboratory features, as well as complications and the treatment of tuberculous otitis media.

Study on the correlation between OSAS and thoracic deformity in children: A retrospective single-center study in China.

OBJECTIVE: To evaluate the correlation between obstructive sleep apnea syndrome (OSAS) and the development of thoracic deformity in Children. METHODS: A retrospective analysis was performed with the medical records of 39 pediatric OSAS patients with thoracic deformity and matching 39 without thoracic deformity as control group between January 2015 and June 2019. The contrast was performed with age, gender, height, weight, body mass index (BMI), apnea/hypopnea index (AHI), the lowest oxyhemoglobin saturation (loSpO2)at night, tonsil and adenoid size, Alkaline phosphatase (ALP)and trace elements and metals between two groups. RESULTS: BMI, AHI, the lowest SpO2, Phosphorus and Zinc were the risk factors of thoracic deformity. Age, gender, disease history, the size of tonsil and adenoid, ALP and other trace elements were no significant difference occurred between two groups. CONCLUSION: OSAS characterized by apnea and hypoxia which are caused by narrow upper airway may be one cause of thoracic deformity in children. Pediatricians, thoracic and otolaryngologic surgeons should be alert to OSAS when thoracic deformities are diagnosed in children.

Is there "hidden hearing loss" in patients with chronic rhinosinusitis?

Objective: This study was to investigate whether there is impairment of auditory function in chronic rhinosinusitis (CRS).Study sample: A total of 85 patients were allocated into either the CRS group (n = 65) or a simple deviated nasal septum group (n = 20). Both groups without other risk factors for sensorineural hearing loss exhibited normal thresholds at standard audiometric frequencies. Another group (n = 30) of healthy subjects without CRS or a deviated nasal septum were gender and age matched.Design: Analyse the results of audiology test including pure tone audiometry, an acoustic impedance test, distortion product otoacoustic emissions (DPOAE) and the auditory brainstem response (ABR) for each subject analyse the test results of for each object.Results: The group differences were statistically significant for each high-frequency pure tone (p < 0.05). The ABR showed a difference between groups in amplitude. The DPOAE pass rate of the CRS group was lower than that of the control group.Conclusions: This study showed a significant correlation between CRS and auditory impairment. CRS might impair cochlear functions by damaging inner ear hair cells and/or, outer hair cells (OHCs), consequently altering the activity of the entire auditory pathway originating in the ventral cochlear nucleus (VCN) to the inferior colliculus.

Reversal effect of FW-04-806, a macrolide dilactone compound, on multidrug resistance mediated by ABCB1 and ABCG2 in vitro and in vivo.

BACKGROUND: Overexpression of ATP-binding cassette (ABC) transporters, such as ABCB1 and ABCG2, has been proved to be a major trigger for multidrug resistance (MDR) in certain types of cancer. A promising approach to reverse MDR is the combined use of nontoxic and potent ABC transporters inhibitor with conventional anticancer drugs. We previously reported that FW-04-806 (conglobatin) as a novel Hsp90 inhibitor with low toxicity, capable of attenuating Hsp90/Cdc37 /clients interactions and producing antitumor action in vitro and in vivo. Our early activity screening found that FW-04-806 at non-cytotoxic concentration was able to enhance the cytotoxicity of chemotherapeutic agents on the ABCB1 overexpressing cells. Therefore, we speculated that FW-04-806 might be a promising MDR reversal agent. In the present study we further investigated its reversal effect of MDR induced by ABC transporters in vitro and in vivo. METHODS: MTT assay in vitro and xenograftes in vivo were used to investigate reversal effect of FW-04-806 on MDR in ABCB1 or ABCG2 overexpressing cancer cells. To understand the mechanisms for the MDR reversal, we examined the effects of FW-04-806 on intracellular accumulation of doxorubicin (DOX, adriamycin, adr)/Rhodamine 123 (Rho 123), efflux of doxorubicin, expression levels of gene and protein of ABCB1 or ABCG2 and ATPase activity of ABCB1, and carried out molecular docking between FW-04-806 and human ABCB1. RESULTS: The results indicated that FW-04-806 significantly enhanced the cytotoxicity of substrate chemotherapeutic agents on the ABCB1 or ABCG2 overexpressing cells in vitro and in vivo suggesting its reversal MDR effects. FW-04-806 increased the intracellular accumulation of DOX or Rho123 by inhibiting the efflux function of ABC transporters in MDR cells rather than in their parental sensitive cells. However, unlike other ABC transporter inhibitors, FW-04-806 had no effect on the ATPase activity nor on the expression of ABCB1 or ABCG2 on either mRNA or protein level. Molecular docking suggested that FW-04-806 may have lower affinity to the ATPase site, which was consistent with its no significant effect on the ATPase activity of ABCB1; However FW-04-806 may bind to substrate binding site in TMDs more stably than substrate anticancer drugs therefore obstruct the anticancer drugs pumped out of the cell. CONCLUSIONS: FW-04-806 is a compound that has both anti-tumor and reversal MDR effects, and its antitumor clinical application is worth further study.

Study on the Clinical Features and Prognosis of Penicilliosis marneffei Without Human Immunodeficiency Virus Infection.

OBJECTIVE: To improve the diagnosis and treatment of Penicilliosis marneffei without human immunodeficiency virus infection. METHODS: Analyze and review the clinical features, diagnosis and treatment of six cases of P. marneffei without human immunodeficiency virus infection at The First Affiliated Hospital of Fujian Medical University. RESULTS: Two cases were diagnosed in the ENT Department, three cases in the respiratory department and one case in the dermatological department. Penicillium marneffei infection was confirmed by sputum culture, blood culture and tissue biopsy. After definite diagnosis, one refused further treatment, and others showed significant improvement. CONCLUSION: Penicilliosis marneffei is insidious onset and easy to be escaped and misdiagnosed. To achieve early diagnosis and appropriate treatment, doubtful cases should be alerted for the diagnoses as P. marneffei.