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A rapid, on-site, and accurate SARS-CoV-2 detection method is crucial for the prevention and control of the COVID-19 epidemic. However, such an ideal screening technology has not yet been developed for the diagnosis of SARS-CoV-2. Here, we have developed a deep learning-based surface-enhanced Raman spectroscopy technique for the sensitive, rapid, and on-site detection of the SARS-CoV-2 antigen in the throat swabs or sputum from 30 confirmed COVID-19 patients. A Raman database based on the spike protein of SARS-CoV-2 was established from experiments and theoretical calculations. The corresponding biochemical foundation for this method is also discussed. The deep learning model could predict the SARS-CoV-2 antigen with an identification accuracy of 87.7%. These results suggested that this method has great potential for the diagnosis, monitoring, and control of SARS-CoV-2 worldwide.

The current study was performed to investigate the potential association of serum CXCL12 with disease severity in non-traumatic ONFH.This study enrolled 182 patients with non-traumatic ONFH and 182 age- and gender-matched healthy controls. The CXCL12 levels in serum were measured by enzyme-linked immunosorbent assay. Meanwhile, serum levels of procollagen type I (PINP) and Interleukin-33(IL-33) were also detected. The radiographic severity was determined by FICAT grade. Clinical severity was evaluated by visual analogue scale (VAS), Harris Hip Score (HHS) and Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC). Among the non-traumatic ONFH, 90 patients ONFH received total hip arthroplasty, the localization and expression of the CXCL12 protein and mRNA were detected by immunohistochemistry, Western blot analysis, RT-PCR and in necrotic area and adjacent non-necrotic area from lesioned femoral neck from ONFH patients and healthy femoral head from femoral neck fracture patients. Receiver operating characteristic (ROC) curve analysis was carried out to confirm the diagnostic value serum CXCL12, PINP and IL-33 with regard to the FICAT grade.Serum CXCL12 levels were significantly lower in non-traumatic ONFH patients compared with healthy controls. CXCL12 mRNA and protein expressions were both significantly decreased in necrotic area in comparison with non-necrotic area and healthy femoral head. Serum CXCL12 concentrations were drastically reduced in patients with FICAT stage 4 compared with stage 3, and CXCL12 concentrations in patients with stage 3 were markedly lower than stage 2. Serum CXCL12 levels were negatively related to FICAT grading. In addition, Serum CXCL12 concentrations were also negatively related to VAS, WOMAC scores and positively correlated with HHS scores. Meanwhile, serum CXCL12 levels were positively correlated with serum PINP and negatively correlated with IL-33 levels. ROC curve analysis implicated that decrease CXCL12 in serum may act as a favorable marker for FICAT grade.Decreased serum CXCL12 concentrations may reflect disease severity of non-traumatic ONFH.

Understanding viral tropism is an essential step toward reducing severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) transmission, decreasing mortality from coronavirus disease 2019 (COVID-19) and limiting opportunities for mutant strains to arise. Currently, little is known about the extent to which distinct tissue sites in the human head and neck region and proximal respiratory tract selectively permit SARS-CoV-2 infection and replication. In this translational study, we discover key variabilities in expression of angiotensin-converting enzyme 2 (ACE2) and transmembrane serine protease 2 (TMPRSS2), essential SARS-CoV-2 entry factors, among the mucosal tissues of the human proximal airways. We show that SARS-CoV-2 infection is present in all examined head and neck tissues, with a notable tropism for the nasal cavity and tracheal mucosa. Finally, we uncover an association between smoking and higher SARS-CoV-2 viral infection in the human proximal airway, which may explain the increased susceptibility of smokers to developing severe COVID-19. This is at least partially explained by differences in interferon (IFN)-β1 levels between smokers and non-smokers., Graphical Abstract, Nakayama et al. discover key variabilities in expression of SARS-CoV-2 entry factors among human head and neck mucosal tissues. They demonstrate that SARS-CoV-2 preferentially infects the nasal cavity and trachea. They uncover an association between smoking and higher SARS-CoV-2 in the human proximal airway, in part because of differences in IFN-β1 expression.

Emerging evidence suggests that chronic rhinosinusitis with nasal polyps (CRSwNP) is a highly heterogeneous disease with disparate inflammatory characteristics between different racial groups and geographies. Currently, little is known about possible underlying distinguishing factors between these inflammatory differences.Our aim was to interrogate differences in CRSwNP disease between White/non-Asian patients and Japanese patients by using whole transcriptome and single-cell RNA gene expression profiling of nasal polyps (NPs).We performed whole transcriptome RNA sequencing with endotype stratification of NPs from 8 White patients (residing in the United States) and 9 Japanese patients (residing in Japan). Reproducibility was confirmed by quantitative PCR in an independent validation set of 46 White and 31 Japanese patients. Single-cell RNA sequencing (scRNAseq) was used to stratify key cell types for contributory transcriptional signatures.Unsupervised clustering analysis identified 2 major endotypes that were present within both cohorts of patients with NPs and had previously been reported at the cytokine level: (1) type 2 endotype and (2) non-type 2 endotype. Importantly, there was a statistically significant difference in the proportion of these endotypes between these geographically distinct subgroups with NPs (P = .03). Droplet-based single-cell RNA sequencing further identified prominent type 2 inflammatory transcript expression: C-C motif chemokine ligand 13 (CCL13) and CCL18 in M2 macrophages, as well as cystatin SN (CST1) and CCL26 in basal, suprabasal, and secretory epithelial cells.NPs from both racial groups harbor the same 2 major endotypes, which we have determined to be present in differing ratios between each cohort with CRSwNP disease. Distinct inflammatory and epithelial cells contribute to the type 2 inflammatory profiles observed.

Background Increasing evidence has suggested that autophagy may play a resistant role during photodynamic therapy (PDT). The Wnt/β-catenin pathway is tightly involved in cell proliferation and autophagy. In this study, we aimed to determine the influence of 5-Ethylamino-9-diethylaminobenzo[a]phenoselenazinium (EtNBSe) mediated PDT (EtNBSe-PDT) on autophagy, proliferation and Wnt/β-catenin pathway in human NPC cell line (HNE-1 cells), and further explore the underlying crosstalk between them. Methods Cell viability and proliferation was evaluated by MTT assay. Autophagy and Wnt/β-catenin signaling pathway was analyzed by western blotting and immunofluorescence. Results It was revealed that EtNBSe-PDT significantly impeded the viability and proliferation of HNE-1 cells. Meanwhile EtNBSe-PDT could notably induce autophagy in HNE-1 cells accompanied with the inhibition of Wnt/β-catenin pathway. The Wnt/β-catenin pathway activator Wnt agonist was found to partially counteract the inhibitory proliferation of HNE-1 cells and suppress the autophagy induced by EtNBSe-PDT. In addition, pretreatment with the autophagy inhibitor 3-methyladenine (3-MA) or Wnt agonist showed the potential in enhancing the cytotoxic effect of EtNBSe-PDT (cell survival from 50.71 ± 4.16% to 24.53 ± 4.27% and from 52.64 ± 3.54% to 35.74 ± 4.27% respectively). Conclusion Taken together, this study demonstrated that EtNBSe-PDT suppressed viability and proliferation, and induced autophagy of HNE-1 cells via downregulating the Wnt/β-catenin pathway. The autophagy further constituted the cytoprotective mechanisms involved in HNE-1 cells, which suggested that the combination of EtNBSe-PDT and autophagy inhibitors may be a promising strategy for the treatment of human NPC.

Long noncoding RNAs (lncRNAs) CASC11 is an oncogenic lncRNA in gastric cancer and colorectal cancer. Our study aimed to investigate the role of lncRNA CASC11 in bladder cancer. In this study we showed that plasma lncRNA CASC11 was upregulated, while plasma miRNA‐150 was downregulated in patients with early‐stage bladder cancer than in healthy controls. Altered expression of plasma lncRNA CASC11 and miRNA‐150 separated patients with bladder cancer from healthy controls. lncRNA CASC11 expression was inversely correlated with miRNA‐150 expression in patients with bladder cance but not in healthy controls. Overexpression of lncRNA CASC11 mediated the inhibition of miRNA‐150 expression in cancer cells, while miRNA‐150 overexpression did not significantly alter lncRNA CASC11 expression. lncRNA CASC11 overexpression promoted, while miRNA‐150 overexpression inhibited cancer cell proliferation. miRNA‐150 also attenuated the enhancing effects of lncRNA CASC11 overexpression on cancer cell proliferation. However, overexpression of lncRNA CASC11 showed no significant effects on cancer cell migration and invasion. Therefore, lncRNA CASC11 may promote cancer cell proliferation in bladder cancer, and the actions of lncRNA CASC11 are likely through miRNA‐150.

Androgens are essential for prostate cancer development. However, steroidogenesis has mainly been investigated in a limited number of prostate cancer cell lines, leading to varied conclusions and elusive clinical significance. Here, we established an ex vivo research platform with fresh biopsy samples transiently cultured with tritium- labelled androgens to trace steroidogenesis in prostate tissues and investigate its potential clinical application. DHEA was confirmed as the major precursor for androgen synthesis in the prostate. Significant amounts of oxidized DHEA and 5α-androstanedione were generated from DHEA in prostate biopsy samples. Prostatic steroidogenesis was independent of other clinical factors. Furthermore, prostatic steroidogenesis was suppressed after androgen deprivation therapy but increased upon treatment resistance, indicating that prostatic steroidogenesis was affected by clinical treatments. Overall, we provide an accessible research platform to characterize steroidogenesis in prostate tissue and indicate the correlation between prostatic steroidogenesis and disease progression.

The edible and medicinal part of Inula nervosa Wall. (Xiaoheiyao) is confined to its root without sufficient phytochemical and biological investigation. In this study, the secondary metabolites of root, stem, leaf, and flower of I. nervosa Wall. were visualized using Global Natural Products Social Molecular Networking (GNPS), MolNetEnhancer, XCMS(xcmsonline.scripps.edu) analysis, and `ili mapping based on high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) data to reveal their chemical differences. Among the 11 kinds of chemical repertoires annotated by MolNetEnhancer and 16 hits against the GNPS library, 10-isobutyryloxy-8,9-epoxythymol isobutyrate (1) was revealed as the most dominant and responsible marker between the roots and the other parts. Moreover, a battery of unique MS features as well as differential markers were discovered from different parts of the plant. The chemical differences contribute to the bioactivity differences, which presented in the 2,2-diphenyl-1-picryl-hydrazyl (DPPH)assay and H2O2-insulted HepG2 cells and were in significant correlations with the contents of 1. real-time reverse transcription polymerase chain reaction (RT-PCR)results demonstrated that I. nervosa Wall. extracts upregulated the mRNA expression of nuclear factor E2-related factor 2(Nrf2), heme oxygenase 1(HO-1), NAD(P)H quinone dehydrogenase 1 (NQO1), manganese superoxide dismutase (MnSOD), and glutamate-cysteine ligase catalytic subunit (GCLC) actors involved in antioxidative response in H2O2-challenged HepG2 cells. These findings support the roots of I. nervosa Wall. as active parts of Xiaoheiyao, and also indicate the potential antioxidant activities of other parts.

Online healthcare services are growing rapidly. Patients can receive health support through communication with online health professionals. However, previous research on eHealth has focused on patients&rsquo, online behavioral intentions. The purpose of this study is to investigate the effect of online patient&ndash, doctor communication on offline behavioral intentions and to propose a conceptual model for eHealth. We integrate fairness theory and band extension theory to develop a research model. This is further tested by structural equation modeling (SEM), with 279 valid sets of data from patients on the online health platform. Using partial least squares analysis (PLS), the results show that perceived fairness is an important factor affecting online satisfaction and the willingness to ask online doctors, which in turn has a positive impact on the willingness to go to offline doctors. These findings shed light on the research model for eHealth and offer suggestions for improving patients&rsquo, online satisfaction, as well as both online and offline behavioral intentions.

BACKGROUND Leptocarpin (LTC) has drawn much attention for suppressing tumor growth or reducing inflammation. However, the effect of LTC on osteosarcoma has rarely been reported. Our object was to determine whether LTC suppresses MG63 cell proliferation, migration, and invasion, and whether type-1 insulin-like growth factor receptor (IGF-1R) is one of the targets in LTC suppressing osteosarcoma. MATERIAL AND METHODS Cytotoxicity of LTC was performed by use of a cell-counting kit-8 (CCK-8). RNA interference (RNAi) or pEABE-bleo IGF-1R plasmid were used for silencing or overexpressing IGF-1R, Western blot (WB) analysis was used for IGF-1R expression, CCK-8 for proliferation, and transwell assay for migration and invasion. RESULTS LTC (23.533 μM) treatment for 48 h was taken as the 50% inhibiting concentration (IC50), which significantly (P

We investigated the relative ratios of collagen composition of periarticular tissue of the elbow, wrist, metacarpophalangeal, proximal and distal interphalangeal joints. Periarticulat tissue, which we defined as the ligaments, palmar plate and capsule, was harvested from ten fresh-frozen cadaveric upper limbs, yielding 50 samples. The mean paired differences (95% confidence interval) of the relative ratios of collagen between the five different joints were estimated using mRNA expression of collagen in the periarticular tissue. We found that the relative collagen composition of the elbow was not significantly different to that of the proximal interphalangeal joint, nor between the proximal interphalangeal joint and distal interphalangeal joint, whereas the differences in collagen composition between all the other paired comparisons of the joints had confidence intervals that did not include zero.

Clear cell renal cell carcinoma (ccRCC) has been associated with one of the highest mortality rates among all cancers. Fatty acid binding proteins (FABPs) are 14‑15 kDa proteins that are highly abundant in the cytosol of most tissues. FABP5, a member of the FABP family, has been observed to promote tumor cell growth in numerous cancer types. In order to investigate the function of FABP5 in ccRCC cells in the present study, RNA sequencing data from The Cancer Genome Atlas were analyzed to determine the expression levels of FABP5 in ccRCC patient samples. Survival and Cox regression analyses were performed to measure the association between FABP5 expression and clinicopathological features of patients with ccRCC. Subsequent in vitro experiments downregulated or overexpressed FABP5 in Caki‑1 and 786O ccRCC cells using lentiviral vectors to evaluate cell proliferation ability, and a xenograft transplantation model was established to examine the effect of FABP5 on tumorigenesis in vivo. The results demonstrated that FABP5 expression was significantly upregulated in samples from patients with ccRCC when compared with normal tissue samples. High FABP5 expression was also significantly correlated with tumor and metastasis classifications and predicted poor survival in patients with ccRCC. In ccRCC cells, silencing of FABP5 significantly inhibited cell proliferation, while overexpression of FABP5 promoted cell proliferation when compared to the respective controls. In addition, treatment with the phosphatidylinositol‑4,5‑bisphosphate 3‑kinase (PI3K)/AKT inhibitor, LY294002, attenuated the pro‑proliferative effects of exogenous FABP5 expression in Caki‑1 and 786O cells. This indicated that the PI3K/AKT signaling pathway may be partially involved in the FABP5‑mediated increase in ccRCC cell proliferation. Furthermore, FABP5 was observed to regulate tumor growth in nude mice in vivo. In conclusion, the results of the present study suggest that FABP5 may exert a pro‑proliferative role in ccRCC and may be associated with malignant progression and tumorigenesis.

Background Mannose-binding lectin (MBL) is a key component of innate immunity. The expression of cortical MBL is up-regulated after clinical and experimental head trauma. This study aimed to assess the association of serum MBL levels with injury severity and long-term clinical outcomes after severe traumatic brain injury (STBI). Methods Serum MBL levels were measured in 122 patients and 100 healthy controls. Multivariate analyses were used to analyze the relationship between serum MBL levels and trauma severity reflected by Glasgow Coma Scale scores as well as between serum MBL levels and 6-month mortality and unfavorable outcome (Glasgow Outcome Scale score: 1–3). A receiver operating characteristic (ROC) curve was structured to evaluate the prognostic predictive performance of serum MBL levels. Results Compared with healthy controls, serum MBL levels of patients were markedly elevated. Using multivariate analyses, serum MBL levels were found to be associated closely with Glasgow Coma Scale (GCS) scores and MBL emerged as an independent predictor for 6-month mortality and unfavorable outcome. Under ROC curve, serum MBL levels and GCS scores possessed similar prognostic predictive values. Conclusion Increased serum level of MBL was independently associated with head trauma severity and long-term clinical outcomes of STBI.

Background: To investigate the effect of myostatin (GDF-8) stimulation of bone marrow derived mesenchymal stem cells (BMSCs) on tenogenesis in the setting of tendon repair. GDF-8 has demonstrated the ability to augment tenogenesis and we sought to identify if this effect could lead to the focused differentiation of pluripotential stem cells down a tenocyte lineage ex vivo.Methods: Cadaveric upper limb flexor tendons were harvested, decellularized and divided into 1 cm segments. Sutures seeded with stem cells were passed through tendon segments to simulate repair. The repaired tendons were then cultured either with or without myostatin for 3, 5, and 7 days. The experiment was also repeated with non-decellularized tendons for a total of 4 groups. The tendons were then evaluated for the expression of scleraxis and tenomodulin, two biomarkers for tendon.Results: Myostatin stimulation led to an increase in expression of tenomodulin and scleraxis at 5 and 7 days in both the decellularized and non-decellularized tendons. Myostatin increased the differentiation of BMSCs into tenocytes and/or led to the upregulation of tenomodulin and scleraxis production by the native tenocytes present within the non-decellularized tendons.Conclusions: The addition of myostatin to BMSCs leads to tenocyte differentiation as evidenced by the expression of tenocyte biomarkers, scleraxis and tenomodulin. This effect is maintained in an ex vivo tendon repair model suggestive that these cells survive the passage through tendon tissue and remain metabolically active.

Natural killer (NK) cells are cytotoxic type 1 innate lymphoid cells (ILCs) that defend against viruses and mediate anti-tumor responses, yet mechanisms controlling their development and function remain incompletely understood. We hypothesized that the abundantly expressed microRNA-142 (miR-142) is a critical regulator of type 1 ILC biology. Interleukin-15 (IL-15) signaling induced miR-142 expression, whereas global and ILC-specific miR-142-deficient mice exhibited a cell-intrinsic loss of NK cells. Death of NK cells resulted from diminished IL-15 receptor signaling within miR-142-deficient mice, likely via reduced suppressor of cytokine signaling-1 (Socs1) regulation by miR-142-5p. ILCs persisting in Mir142-/- mice demonstrated increased expression of the miR-142-3p target αV integrin, which supported their survival. Global miR-142-deficient mice exhibited an expansion of ILC1-like cells concurrent with increased transforming growth factor-β (TGF-β) signaling. Further, miR-142-deficient mice had reduced NK-cell-dependent function and increased susceptibility to murine cytomegalovirus (MCMV) infection. Thus, miR-142 critically integrates environmental cues for proper type 1 ILC homeostasis and defense against viral infection.

Severe forms of chronic rhinosinusitis (CRS), a common upper airway inflammatory disorder, are associated with nasal polyps (NPs). NP disease is ameliorated by glucocorticoid (GC) treatment, whose cellular effects are poorly understood. We therefore assessed the influence of GC therapy on NPs in CRS patients, focusing on regulatory T (Treg) cells. Treg cell populations were analyzed by flow cytometry in NPs and control tissues from GC-treated CRS patients and controls. After GC exposure, selective expansion of Treg cells was seen within NPs, and not blood or adjacent ethmoid tissues. To confirm direct GC effects, NPs from the same patients were biopsied prior to, and following, 1week of oral GC exposure. Direct expansion of Tregs into the same NP bed was detected in 4/4 CRS patients following GC exposure. Treg cell spikes into NPs were secondary to cellular recruitment given limited Ki67 expression within these regulatory cells. Chemokine gene expression profiling identified several chemokines, notably CCL4, induced within NPs upon GC treatment. Neutralization of chemokine receptor/ligand interactions using CCR4 small molecule antagonists reduced Treg migration towards GC-treated NPs in an ex vivo migration assay. Our findings suggest that the common use of GCs in the treatment of NP disease leads to recruitment of Treg cells from peripheral sites into NP tissues, which may be critical to the anti-inflammatory effect of GCs. Mechanistically Treg expansion appears to be conferred, in part, by chemokine receptor/ligand interactions induced following corticosteroid therapy.

Purpose To describe histologic evidence of nerve trauma during the creation and use of the 3-4 portal. Methods Fourteen fresh-frozen cadaveric wrists were mounted on a custom-built frame that simulated a wrist arthroscopy traction tower. After the 3-4 portal was created in the usual manner, the skin was dissected off to identify possible trauma to the posterior interosseous nerve (PIN). Specimens were categorized into those where there was clearly no trauma to the PIN and those where trauma was possible. In the cases where trauma was possible, we harvested the PIN with a cuff of the proximal edge of the portal and examined the cross-sectional histology of the most distal sections for the presence of neural tissue. Results There was clearly no trauma to the PIN in 3 of the wrists during the creation of the 3-4 portal. In the remaining 11 wrists with possible trauma to the PIN, we identified axonal tissue on histologic examination at the proximal edge of the 3-4 portal in 7 of these specimens. In summary, 50% (7 of 14) of our specimens had visual and histologic evidence of trauma to the PIN. Conclusions Based on the findings of this study, there may be more instances of trauma to the PIN during routine wrist arthroscopy than have been previously reported. Clinical Relevance Findings suggest that transection or injury to this nerve may not lead to any clinical sequelae. However, if there is an instance where a patient has persistent, otherwise unexplained, dorsal wrist pain after a wrist arthroscopy procedure, iatrogenic neuroma of the PIN may be responsible and should be considered.

This study aims to investigate the effect of non-transecting anastomotic urethroplasty for treatment of posterior urethral stricture. A total of 23 patients with traumatic posterior urethral stricture were enrolled and then divided into two groups. In one group, 12 patients underwent non-transecting anastomotic urethroplasty. In the other group, 11 patients underwent conventional posterior urethra end-to-end anastomosis. The effect of operation was evaluated using the following parameters: the bleeding amount during operation, operation time, IIEF-5 scores after operation, maximum flow rate (Qmax), and rating scale of quality of life (QoL). The comparison between the conventional posterior urethra end-to-end anastomosis group and the non-transecting anastomotic urethroplasty group showed no significant difference with regard to average operation time. However, a significant difference was observed between the groups with regard to the bleeding amount during operation. The patients in the group of non-transecting anastomotic urethroplasty urinated smoothly after the removal of catheter. Meanwhile, one patient from the group of conventional posterior urethra end-to-end anastomosis had difficulty urinating after the removal of catheter. Furthermore, significant differences in the operation time, bleeding amount during operation, IIEF-5 scores after operation, and rating scale of QoL were observed, whereas no significant difference was observed between urine flow rates of the two groups after operation. Overall, nontransecting anastomotic urethroplasty is effective for posterior urethra reconstruction, and it can reduce the occurrence rate of erectile dysfunction after operation.

Thioredoxin (TRX), a potent anti-oxidant, is released during inflammation and oxidative stress. The purpose of this study was to establish the relationship between serum TRX concentrations and trauma severity and outcome in severe traumatic brain injury (STBI).We determined serum TRX concentrations in 112 patients and 112 controls. Multivariate analyses were performed to analyze the predictive factors of 1-week mortality, 6-month mortality and 6-month unfavorable outcome. The predictive values were investigated under receiver operating characteristic curves.Serum TRX concentrations were markedly higher in patients than in controls (19.1±7.8ng/ml vs. 8.0±2.3ng/ml, P0.001). There was a significant negative association between serum TRX concentrations and Glasgow coma scale (GCS) scores (r=-0.543, P0.001). Increased TRX was identified as an independent prognostic marker of 1-week mortality [Odds ratio (OR), 1.220; 95% confidence interval (CI), 1.101-1.367; P0.001], 6-month mortality (OR, 1.201; 95% CI, 1.097-1.324; P0.001) and 6-month unfavorable outcome (OR, 1.189; 95% CI, 1.090-1.311; P0.001). TRX concentrations improved area under curve of GCS scores for 6-month unfavorable outcome, but not for 1-week mortality and 6-month mortality.Increased serum TRX concentration, associated highly with trauma severity and poor outcome, might be a novel prognostic marker in patients with STBI.

To determine if statins induce anti-inflammatory effects in upper airway inflammation. Mediators of innate and adaptive immunity regulate airway inflammation. Release of these mediators involves enzymatic conversion of polyunsaturated fatty acids into biologically active mediators, which can be blocked by statins. Although upper airway inflammation and chronic sinusitis occur in millions of patients with asthma worldwide, the anti-inflammatory effects of statins in upper airway inflammation have not been previously studied.Laboratory research.Tertiary referral center.Analysis of sinus tissues collected from patients with chronic rhinosinusitis revealed suppression of highly expressed inflammatory mediators in patients who were found to be on statins, suggesting that statins may induce anti-inflammatory effects. Therefore, the authors performed an in vitro study to determine if these anti-inflammatory effects were induced by statins. Cultured primary human airway epithelial cells were exposed to ambient air pollution particulates (PM) to trigger the inflammation, with and without statins, and the expression of inflammatory mediators was analyzed.The authors found that expression of CCL5, CCL11, and IL13RA was suppressed in patients on statins. In vitro exposure to PM enhanced the expression of these mediators, while pretreatment with statins completely blocked these effects. Furthermore, the effects of statins were blocked by inhibition of the statin pathway using isopentenyl-5-pyrophosphate. Statins did not have any significant effect on the viability of normal cells.Statins induce anti-inflammatory effects in human airway epithelial inflammation. Statins may play a role in the treatment and prevention of chronic rhinosinusitis and pulmonary exacerbation of obstructive airway diseases.

Supplemental Digital Content is available in the text, Osteoporosis or osteopenia is a common complication in patients with cirrhosis, but little is known about the risk factors for the occurrence of osteoporosis. Patients with liver cirrhosis due to chronic virus infection and alcoholic abuse were enrolled. Bone mineral density (BMD) was determined using dual-energy x-ray absorptiometry (DXA). Osteoporosis was diagnosed according to WHO criteria. The severity of liver stiffness was measured by Fibroscan. Demographic data, such as age, gender, weight, height, and body mass index (BMI), were collected. Logistic regression analysis was used to recognize the risk factors of osteoporosis in patients with cirrhosis. A total of 446 patients were included in this study: 217 had liver cirrhosis (male, 74.2%; mean age, 57.2 ± 10.27) and 229 were matched controls (male, 69%, mean age, 56.69 ± 9.37). Osteoporosis was found in 44 patients (44/217, 20.3%). The spine and hip BMD in cirrhotic patients were significantly lower than that in controls. When the cirrhotic and control subjects were stratified by age, gender, and BMI, the significant difference was also observed in women patients, patients older than 60, and patients with BMI

Continued smoking causes tumor progression and resistance to therapy in lung cancer. Carcinogens possess the ability to block apoptosis, and thus may induce development of cancers and resistance to therapy. Tobacco carcinogens have been studied widely; however, little is known about the agents that inhibit apoptosis, such as nicotine. We determine whether mitochondrial signaling mediates antiapoptotic effects of nicotine in lung cancer. A549 cells were exposed to nicotine (1 muM) followed by cisplatin (35 muM) plus etoposide (20 muM) for 24 hours. We found that nicotine prevented chemotherapy-induced apoptosis, improved cell survival, and caused modest increases in DNA synthesis. Inhibition of mitogen-activated protein kinase (MAPK) and Akt prevented the antiapoptotic effects of nicotine and decreased chemotherapy-induced apoptosis. Small interfering RNA MAPK kinase-1 blocked antiapoptotic effects of nicotine, whereas small interfering RNA MAPK kinase-2 blocked chemotherapy-induced apoptosis. Nicotine prevented chemotherapy-induced reduction in mitochondrial membrane potential and caspase-9 activation. Antiapoptotic effects of nicotine were blocked by mitochondrial anion channel inhibitor, 4,4'diisothiocyanatostilbene-2,2'disulfonic acid. Chemotherapy enhanced translocation of proapoptotic Bax to the mitochondria, whereas nicotine blocked these effects. Nicotine up-regulated Akt-mediated antiapoptotic X-linked inhibitor of apoptosis protein and phosphorylated proapoptotic Bcl2-antagonist of cell death. The A549-rho0 cells, which lack mitochondrial DNA, demonstrated partial resistance to chemotherapy-induced apoptosis, but blocked the antiapoptotic effects of nicotine. Accordingly, we provide evidence that nicotine modulates mitochondrial signaling and inhibits chemotherapy-induced apoptosis in lung cancer. The mitochondrial regulation of nicotine imposes an important mechanism that can critically impair the treatment of lung cancer, because many cancer-therapeutic agents induce apoptosis via the mitochondrial death pathway. Strategies aimed at understanding nicotine-mediated signaling may facilitate the development of improved therapies in lung cancer.

Overweight and obese males might exhibit a great risk of infertility. However, according to the current studies, the association between elevated male body mass index (BMI) and the clinical adverse results after assisted reproductive technology (ART) remains controversial. Hence, we conducted a meta-analysis to evaluate the effects of raised male BMI on clinical outcomes following ART. PubMed, EMBASE and three Chinese databases were used to identify relevant studies. The primary outcome was clinical pregnancy rate. Secondary outcomes included live birth rate and sperm parameters. A total of 5262 male participants from 10 cohort studies were subjected to meta-analysis. Results indicated that overweight or obese had no significant impact on clinical pregnancy rate [in vitro fertilisation (IVF): odds ratio (OR), 0.73; 95% confidence interval (CI), 0.39-1.39; intracytoplasmic sperm injection (ICSI): OR, 1.03; 95% CI, 0.92-1.15], live birth rate (IVF: OR, 0.91; 95% CI, 0.78-1.06; ICSI: OR, 1.00; 95% CI, 0.50-1.99) and sperm concentration (SMD, -0.28; 95% CI, -0.65 to 0.08) compared with normal weight following IVF/ICSI treatments. Exclusion of any single study and almost all the sensitivity analyses showed that our results were reliable. At present, the role of male BMI in the process of ART is only partly understood and should be further investigated.

Artelon is a degradable biomaterial used for the treatment of osteoarthritis in the carpometacarpal joint of the thumb. The device reportedly works through 2 modes of action-stabilization of the carpometacarpal joint by augmentation of the joint capsule and by formation of a new articular surface at the trapeziometacarpal interface. We present a patient with late failure of arthroscopic hemitrapeziectomy and Artelon interposition that required surgical excision of the Artelon implant and trapeziectomy 4 years postoperatively. Gross and histologic evaluation of the explanted Artelon implant and remaining trapezium revealed lack of articular resurfacing by hyaline ingrowth.

Purpose To evaluate the effect of thermal treatment on neural tissue in the triangular fibrocartilage complex (TFCC), scapholunate interosseous ligament (SLIL), and lunotriquetral interosseous ligament (LTIL). Methods The intact TFCC, SLIL, and LTIL were harvested from cadaveric specimens and treated with a radiofrequency probe as would be performed intraoperatively. Slides were stained using a triple-stain technique for neurotrophin receptor p75, pan-neuronal marker protein gene product 9.5 (PGP 9.5), and 4′,6-diamidino-2-phenylindole for neural identification. Five TFCC, 5 SLIL, and 4 LTIL specimens were imaged with fluorescence microscopy. Imaging software was used to measure fluorescence signals and compare thermally treated areas with adjacent untreated areas. A paired t test was used to compare treated versus untreated areas. P Results For the TFCC, a mean of 94.9% ± 2.7% of PGP 9.5–positive neural tissue was ablated within a mean area of 11.7 ± 2.5 mm 2 ( P = .02). For the SLIL treated from the radiocarpal surface, 97.4% ± 1.0% was ablated to a mean depth of 2.4 ± 0.3 mm from the surface and a mean horizontal spread of 3.4 ± 0.5 mm ( P = .01). For the LTIL, 96.0% ± 1.5% was ablated to a mean depth of 1.7 ± 0.7 mm and a mean horizontal spread of 2.6 ± 1.0 mm ( P = .02). Differences in the presence of neural tissue between treated areas and adjacent untreated areas were statistically significant for all specimens. Conclusions Our study confirms elimination of neuronal markers after thermal treatment of the TFCC, SLIL, and LTIL in cadaveric specimens. This effect penetrates below the surface to innervated collagen tissue that is left structurally intact after treatment. Clinical Relevance Electrothermal treatment as commonly performed to treat symptomatic SLIL, LTIL, and TFCC tears eliminates neuronal tissue in treated areas and may function to relieve pain through a denervation effect.

To observe the effect of electroacupuncture (EA) stimulation of "Fenglong" (ST 40) on blood lipid contents and inflammatory factor levels in hyperlipemia rats so as to elucidate its mechanism underlying improvement of hyperlipemia.Fifty male SD rats were randomized into 5 groups: normal, model, diet -control, EA intervention (EA), diet-control + EA groups, with 10 rats in each group. Hyperlipemia model was established by feeding the animals with high-fat diet for 30 days. After modeling, rats in the diet-control group were fed with routine fodder. EA was applied to bilateral "Fenglong" (ST 40) for 30 min, once daily for 30 days. Following intraperitoneal injection of 1640 culture fluid, the peritoneal fluid was collected and centrifuged for extracting macrophages. Flow cytometry (FCM) was employed to determine the levels of tumor necrosis factor alpha (TNF-alpha) and interleukin-6 (IL-6) after adding fluoresce-labeled antibodies.The contents of serum TC and LDL-C were remarkably higher and HDL-C level was significantly lower in the model group than in the normal group (P0.01). After EA intervention, serum TC and LDL-C showed an apparent decrease (P0.01). Compared with the normal group, percentages of CD11 b, TNF-alpha and IL-6 were significantly increased in the model group (P0.01), while in comparison with the model group, percentages of CD11 b in both EA and diet-control + EA groups, TNF-alpha and IL-6 percentages of macrophages in the diet-control, EA and diet-control + EA groups were notably decreased (P0.05, P0.01). The effects of the diet-control + EA group were considerably superior to those of the diet-control group in lowering CD11 b, TNF-alpha and IL-6 percentages (P0.05, P0.01). No significant differences were found between the diet-control and EA groups in the aforementioned indexes (P0.05). CD11 b level indicates changes of macrophage level due to its specific marker character.EA stimulation of "Fenglong" (ST 40) is effective in lowering serum TC, LDL-C, and macrophage TNF-alpha and IL-6 levels in hyperlipemia rats.

New epithelial cells are generated through the proliferation and differentiation of resident progenitor cells in the nasal cavity. In several upper airway diseases, such as cystic fibrosis and chronic rhinosinusitis, self-renewing progenitor cells may be functionally defective, or compromised in their ability, to regenerate cells that maintain normal mucociliary clearance. Herein, we describe our early work to define and characterize a rare population of human nasal epithelial putative progenitors.Single-cell suspensions of human ethmoid sinus tissues were prepared following endoscopic sinus surgery. Cell surface antibodies were analyzed as candidate markers for detecting progenitor cells. A panel of antibodies, including epithelial cell adhesion molecule (EpCAM, epithelial cells), CD45 (hematopoietic cells), nerve growth factor receptor (NGFR/CD271), intercellular adhesion molecule-1 (ICAM1/CD54), and integrin-α6 (ITGA6/CD49f) were used to resolve epithelial progenitor candidates by high-dimensional flow cytometry and the gating technique of fluorescence minus one (FMO) controls.A rare population of approximately 0.06% of total ethmoid cells was discriminated as EpCAM(-) CD45(-) NGFR(+) ICAM1(+) by surface markers. Use of ITGA6 was excluded based on FMO control analysis. This lineage-negative population was purified to 99% homogeneity by cell sorting and analyzed by immunofluorescence microscopy. Sorted cells were subsequently confirmed to uniformly express the transcription factor p63. Upon in vitro culture, lineage-negative clonal cells were confirmed to spontaneously differentiate into epithelial lineage-positive cells.Using the NGFR and ICAM1 cellular coordinates, we have identified a promising population of native human nasal epithelial progenitor cells that require more formal investigation for their role in upper airway regeneration.

Nasal polyps (NPs) are hallmark inflammatory lesions of sinusitis. Despite the spectrum of NP conditions, cellular differences between NPs from patients with chronic rhinosinusitis with NPs (CRSwNP) and aspirin-exacerbated respiratory disease (AERD) are poorly understood. NPs are associated with abundant eosinophils; the contributions of neutrophil and basophil granulocytes are less defined. We therefore sought to assess granulocyte subpopulations, and differential effects following prednisone pretreatment, within NPs of CRSwNP and AERD patients.NPs, adjacent ethmoid sinus tissue, and peripheral blood mononuclear cells (PBMCs) were obtained from patients undergoing endoscopic sinus surgery. Samples from 5 cohorts: CRSwNP ± prednisone (n = 6 each), AERD ± prednisone (n = 6 each), and controls (n = 9), were analyzed by high-dimensional flow cytometry to gate granulocyte populations. Specimens were also assessed using immunohistochemistry (IHC) staining.Systemic prednisone administration was associated with a lower frequency of eosinophils (p0.0001, n = 6) in NPs in both CRSwNP and AERD patients, whereas a decrease in neutrophils (p = 0.0070, n = 6) in NPs was only observed in CRSwNP patients after prednisone treatment. In contrast, steroids do not alter basophil proportions (p = 0.48, n = 6) within NPs from either group. No significant shift in granulocyte subsets after steroid treatment was identified in the adjacent ethmoid mucosa or PBMCs from the same patients. Immunohistochemistry (IHC) staining supported these findings.Granulocyte subpopulations are focally affected within NPs by systemic steroid exposure, without notable granulocyte alterations in the surrounding regional tissues. These data provide direct insights into the cellular effects of routine prednisone exposure in CRS patients, and highlight a unique microenvironment present within NP lesions.

The airway epithelium generates reactive oxygen species (ROS) as a first line of defense. Dual oxidases (DUOX1 and DUOX2) are the H2 O2 -producing isoforms of the nicotinamide adenine dinucleotide phosphate (NADPH) oxidase family in the airway epithelium. The purpose of this study was to explore the molecular expression, function, and regulation of DUOXs in chronic rhinosinusitis (CRS).Human nasal tissue samples and nasal secretions were collected from 3 groups of patients undergoing sinus surgery (normal, n = 7; CRS with polyposis [CRSwP], n = 6; CRS without polyposis [CRSsP], n = 6). Nasal secretions were studied for cytokine and H2 O2 content. Tissue samples were used to determine DUOX mRNA and protein expression.DUOX1 mRNA level (80.7 ± 60.5) was significantly increased in CRSwP compared to normal (2.7 ± 1.2) and CRSsP (2.3 ± 0.5, p = 0.042). DUOX2 mRNA levels were increased in both CRSwP (18.6 ± 9.9) and CRSsP (4.0 ± 1.3) compared to normal (1.1 ± 0.3; p = 0.008). DUOX protein was found in the apical portion of the nasal epithelium and protein expression was increased in CRSwP and CRSsP. H2 O2 production was significantly higher in CRSwP (160.9 ± 59.4 nM) and CRSsP (81.7 ± 5.6 nM) compared to normal (53.5 ± 11.5 nM, p = 0.032). H2 O2 content of nasal secretions correlated tightly with DUOX expression (p0.001). Cytokines (eotaxin, monokine-induced by interferon γ [MIG], tumor necrosis factor [TNF]-α, interleukin [IL]-8) showed significantly higher levels in nasal secretions from CRSwP compared to normal (p0.05). Levels of eotaxin, MIG, and TNF-α correlated closely with DUOX expression.DUOX1 and DUOX2 were identified as factors upregulated in CRS. Close correlations between DUOX expression and H2 O2 release, and correlation between key inflammatory cytokines and DUOX expression, indicate DUOX in the inflammatory response in CRS.

Somatostatin (SST) may protect organism from overnutrition-induced insulin resistance and oxidative stress by inhibiting pancreatic endocrine and exocrine secretion, gastrointestinal digestion and absorption. Many studies clearly show its release becomes perturbed in diabetes and obesity. Therefore, in the present study we first aimed to investigate whether or not plasma somatostatin level was different in patients with hyperlipidemia and normolipidemic controls. We also assessed the relationship between plasma somatostatin levels with atherosclerotic index (AI) and malondialdehyde (MDA) in non-diabetic dyslipidemic patients. Subjects with hyperlipidemia have insulin resistance and high levels of oxidative stress. Median somatostatin (57.2±19.2 vs 68.0±21.9 pg/mL; p0.05) levels were lower in hyperlipidemic than in normolipidemic subjects. Significant inverse relationships between SST level and AI (r=-0.21, p0.05), or MDA (r=-0.31, p0.01) were observed. These results suggest a possible protective role of endogenous SST, at least on hyperlipidemia and atherosclerosis that are attributed to excess energy intake and physical inactivity. Of course these preliminary results should be supported by prospective studies.

Background Nasopharyngeal cancer (NPC) is a relatively uncommon malignant epithelial cancer seen worldwide. The treatment of NPC has evolved toward combined modality treatment with radiation therapy and chemotherapy. However, chemotherapeutic agents currently have a secondary role, due to their lack of efficacy as curative agents. Recent identification of a novel property of statin drugs raises a promising hope that concurrent use of statins may enhance the efficacy of single-drug chemotherapy in NPC. However, the effects of statins have not been studied before in NPC. In this in vitro study, we demonstrate a unique property of statins that can enhance the efficacy of cisplatin in NPC. Methods Primary human NPC cells (CCL-30) were treated with statins and cisplatin concurrently, and the effects on cell proliferation, apoptosis, and cell colony formation were examined. Results Statins caused significant decrease in cell proliferation and viability in NPC. Statins also induced loss of cell attachment, change in cellular morphology, decrease in colony forming units, and loss of sphere formation in soft gel agar, which are the important properties of tumorigenicity in NPC. Furthermore, we found that the effects of statins occur by a mevalonate (MA)-mediated pathway in these cells. Conclusions We demonstrate a unique property of statins that can enhance the antitumor effects of cisplatin in NPC. Statins may act as a relatively safe and cost effective chemoadjuvant agent in the treatment of NPC. © 2011 ARS-AAOA, LLC.

Background Epidemiological and clinical data implicate that in patients with cancer, continued smoking causes progression of cancer growth and resistance to therapy. The carcinogens possess the ability to block apoptosis, an important mechanism in the development of tumors and resistance to chemotherapy. We previously showed that nicotine enhances growth and proliferation in lung cancer. However, the effects of nicotine, a tobacco carcinogen that inhibits apoptosis, have not been studied before in nasal epithelial carcinoma (NC). In this study, we sought to determine the effects of nicotine on chemotherapy-induced apoptosis in human NC. Methods Primary human NC cells were grown per protocol, treated with combination chemotherapy, and the apoptosis was assessed by TUNEL (terminal deoxynucleotidyl transferase biotin-dUTP nick end labeling) and DNA fragmentation assays. The regulation of mitogen-activated protein kinase (MAPK) and protein kinase B (AKT) signal transduction pathway was examined by real time quantitative polymerized chain reaction, and immunofluorescent staining assays. Results Combination chemotherapy with cisplatin (35 μM) plus etoposide (20 μM) caused a significant increase in NC apoptosis compared with single agent alone, and nicotine, in part, inhibited chemotherapy-induced apoptosis in NC. Furthermore, nicotine induced activation of AKT and MAPK pathways, while inhibition of MAPK using U0126 and AKT by phosphatidylinositol 3-kinase inhibitor, LY294002, in part, blocked the antiapoptotic effects of nicotine against cisplatin and etoposide-induced apoptosis in NC. Conclusion Nicotine inhibits chemotherapy-induced apoptosis in NC via the AKT and MAPK-mediated signaling pathways. We speculate that nicotine may play a role in oncogenesis and resistance to cancer therapy in NC.

Tandem repeat expansion is responsible for the Repeat Expansion Diseases, a group of human genetic disorders that includes Fragile X syndrome (FXS). FXS results from expansion of a premutation (PM) allele having 55-200 CGG.CCG-repeats in the 5' UTR of the FMR1 gene. The mechanism of expansion is unknown. We have treated FX PM mice with potassium bromate (KBrO(3)), a potent DNA oxidizing agent. We then monitored the germline and somatic expansion frequency in the progeny of these animals. We show here that KBrO(3) increased both the level of 8-oxoG in the oocytes of treated animals and the germline expansion frequency. Our data thus suggest that oxidative damage may be a factor that could affect expansion risk in humans.

When using immunocytochemistry, investigators may not know how to optimize staining or how to troubleshoot the method when staining fails. Lacking are guides for comparing techniques and applying information derived from one staining method to another. Newer methods amplify signal detection, but will not necessarily work at the same primary antibody concentrations used for less sensitive reactions. Recommendations of optimal titers are often not accurate and are not usually accompanied by information on the method used to test those antibodies or the specifics of the assay. When the staining does not work, the investigators do not know how to determine if the antiserum is bad, the tissue is bad, or the method is inappropriate for their staining. This unit describes detailed procedures for determining optimal staining and applying that information to three common immunofluorescence methods. Lastly, a formula is provided for converting among the different methods. © 2016 by John WileySons, Inc.

We studied the effects of airborne particulate matters (PM) on cystic fibrosis (CF) epithelium. We noted that PM enhanced human CF bronchial epithelial apoptosis, activated caspase-9 and PARP-1; and reduced mitochondrial membrane potential. Mitochondrial inhibitors (4,4-diisothiocyanatostilbene-2,2′disulfonic acid, rotenone and thenoyltrifluoroacetone) blocked PM-induced generation of reactive oxygen species and apoptosis. PM upregulated pro-apoptotic Bad, Bax, p53 and p21; and enhanced mitochondrial localization of Bax. The anti-apoptotic Bcl-2, Bcl-xl, Mcl-1 and Xiap remained unchanged; however, overexpression of Bcl-xl blocked PM-induced apoptosis. Accordingly, we provide the evidence that PM enhances oxidative stress and mitochondrial signaling mediated apoptosis via the modulation of Bcl family proteins in CF.

Many researches have proved that auditory evoked potentials (AEP) can indicate the depth of anesthesia in the clinical practice. This paper introduces two methods of extracting AEP: the moving time average (MTA) model and the autoregressive model with exogenous input (ARX). In our study we used both the MTA model and the ARX model to extract AEP and the results show that using the ARX model can get the results significantly faster than using the MTA model, and the ARX model is more suitable for the real-time monitoring of the anesthetic depth.

At present, there are at least five kinds of peptide transporters identified in animal tissues, in which PepT1 and PepT2 were discussed most extensively. PepT1 and PepT2 are all members of proton-dependent oligopeptide transporters(POT) family. PepT1 is peptide transporter of low affinity and high capacity whereas PepT2 is high affinity and low capacity. PepT1 is mainly expressed in intestinal tract and weakly expressed in kidney, but PepT2 is mainly expressed in kidney. These transporters have some significant molecular characteristics: (1) Contain 12 membrane-spanning domains and a large extracellular loop between transmembrane domains 9 and 10, and all of the transmembrane domains are highly conserved, the extracellular loops are much less so; (2) The encoded proteins have a number of potential N-glycosylation as well as protein kinase recognition sites, which may be involved in the regulation of peptide transport; (3) His-57 in PepT1 and His-87 in PepT2 are the most critical histidyl residues that are necessary for the uptake function and probably represent some critical binding sites; (4) The proteins range in size from 707 to 729 amino acid residues in various species, with higher homology between species for a given transporter (approximately 80%) and less homology between transporters for a given species (approximately 50%). With the knowledge of the molecular characteristics and tissue distribution of these peptide transporters, we can understand the molecular mechanism underlying the peptide absorption, which may improve the peptide-drugs development in the future.