[The study of smoking impact on autophagy in alveolar macrophages of human silicosis].

Objective: To investigate the effect of smoking on autophagy in alveolar macrophages (AMs) of silicosis patients. Methods: In December 2019, a random sampling method was used to select 42 male patients with silicosis (19 cases of stage II and 23 cases of stage III) who were treated with large volume whole lung lavage from August to December 2017 in the Beidaihe sanatorium. According to the different smoking index of the study subjects (smoking index=smoking cigarette consumptions per day×years of smoking) , we divided them into high (Smoking index>400) , medium (200≤smoking index≤400) , low (smoking index <200) and non-smoking group. The levels of autophagy related proteins LC3, Beclin1, p62 and apoptosis related protein Cleaved Caspase-3 were detected by Western blot. The effects of smoking on autophagy activity of AMs in silicosis were analyzed. Results: The ratio of autophagy related protein LC3 II/LC3 I, the expression of Beclin1, p62, and apoptosis related protein Cleaved Caspase-3 in the high smoking group were significantly higher than that of the middle, low smoking group and the non-smoking group (P<0.05) . Conclusion: Smoking can aggravate the dysfunction of autophagic degradation in silicosis patients' AMs, which may accelerate the progress of silicosis through increasing apoptosis in AMs.

microRNA-144-3p suppresses human neuroblastoma cell proliferation by targeting HOXA7.

OBJECTIVE: Dysregulation of microRNAs (miRNAs) expression often resulted in abnormal cell behaviors. It has been demonstrated that miRs may serve as oncogenic or tumor suppressive functions in tumor. We investigated whether or not miR-144-3p has a role in the progression of human neuroblastoma (NB). PATIENTS AND METHODS: 46 NB patients were enrolled in this study. miR-144-3p expression in NB tissues and cell lines was detected by reverse transcription-quantitative polymerase chain reaction. The biological functions of miR-144-3p in NB were detected by cell counting kit-8 assay, flow cytometry assay, and wound-healing assay. Luciferase activity assay and Western blot assay were performed to validate the direct targets of miR-144-3p. RESULTS: We found miR-144-3p expression was reduced in NB tissues and cell lines and resulted in the stimulation of cell proliferation, cell cycle progression, and cell migration in vitro. Furthermore, we validated homeobox protein A7 (HOXA7) as a direct target of miR-144-3p. CONCLUSIONS: Taken together, these results demonstrated the tumor suppressive role of miR-144-3p in NB and may advance the understanding of the underlying mechanisms of miR-144-3p and HOXA7 in NB.

Bone mineral density as a dose-response predictor for osteoporosis: a propensity score analysis of longitudinal incident study (KCIS no. 39).

BACKGROUND: Osteoporosis is a global disease burden for aging society. The role of quantitative ultrasound (QUS) in the prediction for osteoporosis in a dose-response manner is hardly addressed. AIM: We aimed to show the dose-response of QUS measurement in the prediction for osteoporosis by a community-based study. DESIGN: A prospective cohort study. METHODS: Participants were recruited between 2000 and 2004. Demographic data and heel QUS measurement were collected at baseline. Diagnosis of osteoporosis was ascertained by the follow-up of this cohort over time. In order to reduce the imbalance of baseline characteristics in the observational study, we applied propensity score by using proportional odds regression analysis to match the quintiles of QUS T-score. RESULTS: A total of 44 957 subjects composed of 17 678 men (39.3%) and 27 279 women (69.7%) were recruited. After adjustments for propensity score, an increase in one unit of QUB T-score led to 7% reduction in the risk for osteoporosis [adjusted odds ratio (OR) = 0.93, 95% confidence interval (CI): 0.89-0.96, P < 0.0001]. Higher quintile of QUS T-score yielded a lower risk of osteoporosis with a gradient relationship [OR: 0.82 (95%CI: 0.72-0.92); OR: 0.81 (95%CI: 0.71-0.91); OR: 0.77 (95%CI: 0.68-0.87) and OR: 0.76 (95%CI: 0.67-0.86)] from the second to highest quintile opposed to first quintile (P < 0.0001). The cumulative incidence of osteoporosis was higher in the lower quintile during follow-up (log-rank test, P < 0.001). CONCLUSION: QUS is an independent predictor for osteoporosis in a dose-response manner using a large population-based cohort. Due to the lower cost and portability of QUS measurement, the pre-screening for osteoporosis by QUS can be considered in the area with limited resources can be a feasible and alternative method.

Expressions of miR-525-3p and its target gene SEMG1 in the spermatozoa of patients with asthenozoospermia.

BACKGROUND: Semenogelin 1 (SEMG1) is an important secretory protein in spermatozoa involved in the formation of a gel matrix encasing ejaculated spermatozoa. Previous studies show that the SEMG1 gene is highly expressed in spermatozoa from patients with asthenozoospermia (AZS); however, the underlying molecular mechanisms are not yet clear. OBJECTIVES: To study the molecular mechanism of high expression of SEMG1 gene and its potential roles in AZS. MATERIALS AND METHODS: Western blot and real-time PCR were used to detect the expression levels of SEMG1 protein and mRNA in the ejaculated spermatozoa from normozoospermic males and AZS patients. Bioinformatics analysis was used to predict miRNAs targeting for SEMG1 3'-untranslated region detection of the expression levels of all the candidate miRNAs in ejaculatory spermatozoa in AZS patients or normozoospermic volunteers. Luciferase reporter assays were performed to confirm it can directly bind to SEMG1. Correlation of miR-525-3p and SEMG1 mRNA expression with clinical sperm parameters were also analyzed. Finally, we conducted a follow-up study of reproductive history about all the subjects. RESULTS: SEMG1 mRNA and protein level were significantly higher in AZS patients compared to that in normozoospermic volunteers (p < 0.001). Subsequently, microRNA-525-3p (miR-525-3p) which was predicted as a candidate regulator of SEMG1 was found lower expressed in ejaculatory spermatozoa in AZS patients (p = 0.0074). Luciferase experiment revealed that microRNA-525-3p could directly target SEMG1 3'-untranslated region and suppress its expression. Importantly, our retrospective follow-up study showed that both low miR-525-3p expression and high SEMG1 expression level was significantly associated with low progressive sperm motility, abnormal sperm morphology, and infertility. DISCUSSION AND CONCLUSION: The elevated expression of SEMG1 and reduced expression of miR-525-3p are associated with AZS and male infertility. Our study provides a potential therapeutic target for the treatment of male infertility or for male contraception.

Comprehensive proteomics analysis of exosomes derived from human seminal plasma.

Exosomes are membranous nanovesicles of endocytic origin that carry and transfer regulatory bioactive molecules and mediate intercellular communication between cells and tissues. Although seminal exosomes have been identified in human seminal plasma, their exact composition and possible physiologic function remain unknown. The objective of this study was to perform a comprehensive proteomics analysis of exosomes derived from human seminal plasma. Seminal exosomes were isolated and purified from 12 healthy donors using a 30% sucrose cushion-based exosome-isolation protocol, followed by characterization by western blot, transmission electron microscopy, and nanoparticle tracking analysis before performing extensive liquid chromatography tandem mass spectrometry proteomics analysis. The identified proteins were analyzed by bioinformatics analysis, and seminal exosomes-associated proteins were selectively validated by western blot. A total of 1474 proteins were identified in all seminal exosomes samples, with Gene Ontology analysis demonstrating that these identified seminal exosomes-associated proteins were mostly linked to 'exosomes,' 'cytoplasm,' and 'cytosol.' Bioinformatics analysis indicated that these proteins were mainly involved in biologic processes, including metabolism, energy pathways, protein metabolism, cell growth and maintenance, and transport. Of these identified proteins, PHGDH, LGALS3BP, SEMG1, ACTB, GAPDH, and the exosomal-marker protein ALIX were validated by western blot. This study provided a more comprehensive description of the seminal exosomes proteome and could also be a resource for further screening of biomarkers and comparative proteomics studies, including those associated with male infertility and prostate cancer.

Solute carrier family 12 member 5 promotes tumor invasion/metastasis of bladder urothelial carcinoma by enhancing NF-κB/MMP-7 signaling pathway.

Solute carrier family 12 member 5 (SLC12A5), an integral membrane KCl cotransporter, which maintains chloride homeostasis in neurons, is aberrantly expressed and involved in the tumorigenesis of certain cancers. However, the clinical significance and biological role of SLC12A5 in human bladder urothelial carcinoma (BUC) remains unclear. In this study, the expression of SLC12A5 was examined in clinical specimens of primary BUC and in BUC cell lines using quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR), western blot and immunohistochemistry (IHC). The prognostic value of SLC12A5 expression and its correlation with the clinicopathological features of patients with BUC were analyzed statistically. A series of in vitro and in vivo assays were performed to elucidate the effect of SLC12A5 in BUC and its underlying mechanisms. The present results showed that SLC12A5 expression was significantly increased in BUC tissues. SLC12A5 expression significantly correlated with the tumor node metastasis (TNM) stage. Kaplan-Meier survival curves showed that high SLC12A5 expression was associated with poor survival in patients with BUC. Multivariate analysis indicated that SLC12A5 expression was an independent prognostic marker for the survival of patients. Downregulation of SLC12A5 inhibited the migratory and invasive abilities of BUC cells in vitro, and knocking down SLC12A5 diminished BUC metastasis in vivo. Moreover, we identified that SLC12A5 promoted the migration and invasion of BUC by enhancing MMP-7 expression via NF-κB-dependent transcription. Taken together, our findings indicated that SLC12A5 might function as a tumor metastasis promoting factor in the development and progression of BUC by regulating the NF-κB/MMP-7 signaling pathway. Thus, SLC12A5 might be a prognostic marker as well as a therapeutic target for BUC.

Unmet information needs and clinical characteristics in patients with precancerous oral lesions.

The purpose of this study was to investigate associated factors of the unmet information needs of patients with precancerous oral lesions. For this cross-sectional descriptive study, we recruited patients with precancerous oral lesions from the otolaryngology outpatient department of a single medical centre in central Taiwan. Patients were assessed using a set of structure questionnaires to measure patients' state anxiety levels, attitudes towards cancer prevention and need for information. Patients' anxiety and attitudes towards cancer prevention were evaluated based on unmet needs and associated factors were determined. Among the 106 subjects surveyed, the most prominent unmet information needs were about obtaining the test results as soon as possible. Patients with precancerous oral lesions who had high levels of state anxiety, long duration of time since quitting betel nut chewing and were without a history of oral cancer were more likely to have unmet information needs. A high level of anxiety about precancerous oral lesions was more prevalent among patients with unmet information needs than among those whose information needs were met. Health education and individual counselling should be provided to satisfy the information needs of this population.

Overexpression of p300 correlates with poor prognosis in patients with cutaneous squamous cell carcinoma.

BACKGROUND: It has been suggested that the p300 transcriptional coactivator participates in the regulation of a wide range of cell biological processes, and mutations in p300 have been identified in various cancers. OBJECTIVES: To investigate p300 expression in cutaneous squamous cell carcinoma (cSCC) tissues and its effect on the outcome of patients with cSCC. METHODS: Immunohistochemistry (IHC) was performed on a tissue microarray to investigate p300 expression levels in cSCC tissues. Receiver operating characteristic (ROC) curve analysis, Kaplan-Meier plots and a Cox proportional hazards regression model were used to analyse the data. RESULTS: Based on the ROC curves, we defined the cut-off score for high p300 expression as > 55% of tumour cells positively stained. High expression of p300 was observed in 86 of 165 (52·1%) of the cSCC samples and six of 30 (20%) of the adjacent normal skin tissue samples (P < 0·001). High expression of p300 was positively correlated with lymph node metastasis (P = 0·006) and advanced clinical stage (P < 0·001). In univariate survival analysis, high expression of p300 was correlated with poor patient outcomes in terms of recurrence-free survival (P = 0·006) and overall survival (P < 0·001). Moreover, p300 expression was evaluated as an independent prognostic factor in a multivariate analysis (P = 0·004). CONCLUSIONS: Our results indicate that high p300 expression is associated with aggressive features of cSCC and suggest that p300 expression, as examined by IHC, will be a promising biomarker for predicting clinical outcomes in patients with cSCC.

Low/negative expression of DDX3 might predict poor prognosis in non-smoker patients with oral cancer.

OBJECTIVE: DDX3 has diverse biological functions in translation control, cell growth regulation, and tumor progression. Oral squamous cell carcinoma (OSCC) is a common malignant tumor worldwide with a poor clinical prognosis. The impact of DDX3 expression in OSCC is seldom discussed. MATERIALS AND METHODS: Tumor tissues and adjacent normal tissues were obtained from 324 patients with OSCC. In this study, we used immunohistochemical staining methods to investigate the associations between DDX3 expression and the clinicopathological characteristics of OSCC. RESULTS: Low/negative DDX3 expression in tumor cells was significantly associated OSCC patient characteristics including male gender (P < 0.001), smoking (P < 0.001), alcohol consumption (P < 0.001), betel quid chewing (P = 0.002), poor relapse-free survival (P = 0.001), and poor overall survival (OS) (P = 0.001). Patients with low/negative DDX3 expression, and particularly non-smoker OSCC patients, had significantly worse OS as defined by the log-rank test (P = 0.020 for all cases; P = 0.008 for non-smoker patients). In non-smoker patients with OSCC, low/negative DDX3 expression in tumor cells was associated with poor prognosis (P = 0.024) and a 3.802-fold higher death risk, as determined by Cox regression. CONCLUSIONS: Low/negative DDX3 expression in tumor cells was significantly associated with aggressive clinical manifestations and might be an independent survival predictor, particularly in non-smoker patients with OSCC.

Prediction of lymph node metastasis in T1/T2 tongue squamous cell carcinoma.

OBJECTIVES: In T1, T2, and clinically NO squamous cell carcinoma of the tongue, there is no reliable predictive variable to determine whether or not neck dissection is needed. Thus, we established a predictive score model based on tumour depth and other pathological variables. METHODS: We retrospectively reviewed 115 patients with T1 and T2 stage squamous cell carcinoma of the tongue. Their pathological variables were used to construct a score model for predicting the risk of cervical lymph node metastasis. RESULTS: A predictive score model was proposed using multivariate logistic regression analysis: Score = (2.694 x tumour depth (cm)) + (1.814 x lymphovascular invasion (yes = 1, no = 0)) + (1.175 x perineural invasion (yes = 1, no = 0)). The cutoff point was set at 2.7427. This predictive score model has a sensitivity of 91.2% and specificity of 65.4%. CONCLUSION: A predictive score model was built and a two-stage surgical approach was suggested for T1 and T2 squamous cell carcinoma of the tongue.

Left endobronchial intubation with a double-lumen tube using direct laryngoscopy or the Trachway® video stylet.

We compared direct laryngoscopy with a Macintosh blade vs indirect bronchoscopy with a Trachway® stylet, for endobronchial intubation with a left-sided double-lumen tube. We allocated participants scheduled for thoracic surgery and who had normal predicted laryngoscopy, 30 to each group. The mean (SD) intubation times with laryngoscope and Trachway were 48 (11) s vs 28 (4) s, respectively, p < 0.001. The rates of hoarseness on the first postoperative day, categorised as none/mild/moderate/severe, were 10/12/7/1 and 22/6/2/0, respectively, p = 0.008, without differences on subsequent days. Left endobronchial intubation with a double-lumen tube is slower using direct laryngoscopy and causes more hoarseness than indirect bronchoscopy with a Trachway stylet.

The predictive value of semaphorins 3 expression in biopsies for biochemical recurrence of patients with low- and intermediate-risk prostate cancer.

The class-3 semaphorins (Sema3A-F, Sema3s) are initially identified to play an important role in axonal guidance and cell migration. Our previous studies showed that Sema3s are also involved in the lymph node metastasis of prostate cancer, and are likely to modulate the behavior of prostate cancer with a pro-tumoral or an anti-tumoral effect, depending on their subtypes. However, no study has critically investigated the value of Sema3s expression in preoperative biopsy samples for the prediction of biochemical recurrence (BCR) after radical prostatectomy. In this study, we evaluated Sema3s expression by immunohistochemistry on 198 prostate biopsies with low- and intermediate-risk localized prostate cancer. The median follow-up was 42 months (range, 6-60) for all patients. Our results showed that Sema3A (OR: 0.19, P<0.001), Sema3B (OR: 0.38, P=0.003), Sema3E (OR: 0.39, P=0.007), and Sema3C (OR: 2.31, P=0.014) staining were independent predictors of BCR on multivariable analysis. Sema3A, 3B, 3C and 3E expression demonstrated potential values in predicting BCR upon survival analysis (P=0.001, P=0.003, P=0.029, P=0.037, respectively, Log-rank test). Our findings suggested that Sema3A, 3B, 3C, and 3E immunostaining in prostate biopsies, as supplements to clinicopathological parameters, could be used for predicting BCR in low- and intermediate-risk prostate cancer patients after radical prostatectomy. Specially, concurrent Sema3C-positive and Sema3A-negative, 3B-negative, 3E-negative staining is associated with an adverse prognosis. Further prospective studies in larger patient populations are needed to validate the current observations.

Minimizing shoulder syndrome with intra-operative spinal accessory nerve monitoring for neck dissection.

The objective of this study was to analyze the safety and results of intra-operative SAN (spinal accessary nerve) monitoring during selective neck dissection, with emphasis on shoulder syndrome. Twenty-five consecutive patients with head and neck cancer were studied. Selective neck dissection was performed by a single clinical fellow under the supervision of the department chief using an intra-operative SAN monitor. Electrophysiological data were recorded after initial identification of the SAN and continued until just before closure. Electromyographic evaluation was carried out to assess SAN function one month postoperatively. Shoulder disability was also evaluated at this time using a questionnaire for shoulder syndrome (shrug, flexion, abduction, winging, and pain). No patients had postoperative shoulder syndrome involving shrug, flexion, abduction, or winging. Twenty-two of the 25 (88%) patients had shoulder pain, but the average pain score was low (2.3 ± 1.3). No patients had neck recurrence during at least 1 year of follow up. By using nerve monitoring during selective neck dissection, no patient developed significant "shoulder syndrome", with the exception of slight pain.

Prediction of nodal metastasis in head and neck cancer using a 3T MRI ADC map.

BACKGROUND AND PURPOSE: The detection of cervical nodal metastases is important for the prognosis and treatment of head and neck tumors. The purpose of this study was to assess the ability of ADC values at 3T to distinguish malignant from benign lymph nodes. MATERIALS AND METHODS: From July 2009 to June 2010, twenty-two patients (21 men and 1 woman; mean age, 49.8±9.5 years; age range, 28-66 years) scheduled for surgical treatment of biopsy-proved head and neck cancer were prospectively and consecutively enrolled in this study. All patients were scanned on a 3T imaging unit (Verio) by using a 12-channel head coil combined with a 4-channel neck coil. Histologic findings were the reference standard for the diagnosis of lymph node metastasis. RESULTS: The ADC values derived from the signal intensity averaged across images obtained with b-values of 0 and 800 s/mm2 were 1.086±0.222×10(-3) mm2/s for benign lymph nodes and 0.705±0.118×10(-3) mm2/s for malignant lymph nodes (P<.0001). When an ADC value of 0.851×10(-3) mm2/s was used as a threshold value for differentiating benign from malignant lymph nodes, the best results were obtained with an accuracy of 91.0%, sensitivity of 91.3%, and specificity of 91.1%. CONCLUSIONS: The ADC value is a sensitive and specific parameter that can help to differentiate malignant from benign lymph nodes.

Transnasal endoscopic resection of vidian nerve schwannoma accompanied by sphenoid mucopyocele and oculomotor palsy: a case report.

Schwannomas are rare tumours arising from the peripheral nerve sheath. Nearly half of all schwannomas occur in the head and neck region, but the sinonasal tract is rarely involved. We report on an extremely rare case of vidian nerve schwannoma accompanied by mucopyocele with symptoms of oculomotor palsy and CSF leakage. An exclusively endoscopic endonasal approach was performed to excise the tumour and the dural defect was repaired. To our knowledge, this is the first time a vidian nerve schwannoma has been excised in an exclusively endoscopic approach. We first review the literature and then discuss the benefits for patients undergoing this type of operation.

Survivin SNP-carcinogen interactions in oral cancer.

UNLABELLED: In Taiwan, oral cancer is causally associated with environmental carcinogens. Survivin is an anti-apoptotic protein and is generally considered a marker of malignancy. The current study explored the combined effect of survivin gene polymorphisms and environmental carcinogens on the risk and clinico-pathological development of oral cancer. Five single-nucleotide polymorphisms (SNPs) of survivin genes from 439 male patients with oral cancer and 424 male control participants (who did not have cancer) were analyzed. The survivin -31GG, +9194 GG, and +9809 TT homozygotes exhibited higher risk for oral cancer compared with the corresponding ancestral genotype, after adjustment for related confounders. The survivin -31, +9194, and +9809 SNPs combined with betel quid chewing and/or tobacco consumption could robustly elevate susceptibility to oral cancer. The distribution frequency of the -31 G: +9194 A: +9809 T haplotype was significantly higher in oral cancer patients than in control participants. These results suggest that survivin gene polymorphisms and their interactions with environmental carcinogens may increase susceptibility to oral cancer in Taiwanese men. ABBREVIATIONS: AOR, adjusted odds ratio; CI, confidence intervals; PCR, polymerase chain-reaction; SNP, single-nucleotide polymorphisms.

Endoscope-assisted transoral excision of a huge parapharyngeal pleomorphic adenoma.

The treatment of choice for a parapharyngeal pleomorphic adenoma is total surgical resection. We describe an endoscope-assisted transoral excision of a huge parapharyngeal pleomorphic adenoma, and discuss the benefits of this type of operation.