Ultraspecific One-Pot CRISPR-Based "Green-Yellow-Red" Multiplex Detection Strategy Integrated with Portable Cartridge for Point-of-Care Diagnosis.

Versatile, informative, sensitive, and specific nucleic acid detection plays a crucial role in point-of-care pathogen testing, genotyping, and disease monitoring. In this study, we present a novel one-pot Cas12b-based method coupled with the "Green-Yellow-Red" strategy for multiplex detection. By integrating RT-LAMP amplification and Cas12b cleavage in a single tube, the entire detection process can be completed within 1 h. Our proposed method exhibits high specificity, enabling the discrimination of single-base mutations with detection sensitivity approaching single molecule levels. Additionally, the fluorescent results can be directly observed by the naked eye or automatically analyzed using our custom-designed software Result Analyzer. To realize point-of-care detection, we developed a portable cartridge capable of both heating and fluorescence excitation. In a clinical evaluation involving 20 potentially SARS-CoV-2-infected samples, our method achieved a 100% positive detection rate when compared to standard RT-PCR. Furthermore, the identification of SARS-CoV-2 variants using our method yielded results that were consistent with the sequencing results. Notably, our proposed method demonstrates excellent transferability, allowing for the simultaneous detection of various pathogens and the identification of mutations as low as 0.5% amidst a high background interference. These findings highlight the tremendous potential of our developed method for molecular diagnostics.

Label-Free Colorimetric Method for Detection of Vibrio parahaemolyticus by Trimming the G-Quadruplex DNAzyme with CRISPR/Cas12a.

Vibrio parahaemolyticus (V. parahaemolyticus), which may cause gastrointestinal disorders in humans, is a pathogen commonly found in seafood. There are many methods for detecting V. parahaemolyticus, yet they have some shortcomings, such as high cost, labor-intensiveness, and complicated operation, which are impractical for resource-limited settings. Herein, we present a sequence-specific, label-free, and colorimetric method for visual detection of V. parahaemolyticus. This method utilizes CRISPR/Cas12a to specifically recognize the loop-mediated isothermal amplification (LAMP) products for further trans-cleaving the G-quadruplex DNAzyme and depriving its peroxidase-mimicking activity. In this way, the results can be directly observed with the naked eyes via the color development of 2,2'-azino-di-(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS2-), which displays colorless for positive samples while green for target-free samples. We term such Cas12a-crRNA preventing ABTS2- from developing color by trimming the G-quadruplex DNAzyme as Cascade. The proposed method can detect 9.8 CFU (per reaction) of pure cultured V. parahaemolyticus, and the sensitivity is comparable to real-time LAMP. It has been applied for practical use and showed the capability to detect 6.1 × 102 CFU/mL V. parahaemolyticus in shrimp samples. Based on this, the newly established Cascade method can be employed as a universal biosensing strategy for pathogenic bacterial testing in the field.

Anticancer effect of Limonin against benzo(a)pyrene-induced lung carcinogenesis in Swiss albino mice and the inhibition of A549 cell proliferation through apoptotic pathway.

The main purpose of the current study is to reveal the anticancer action of limonin against benzo(a)pyrene [B(a)P]-treated lung carcinogenesis in Swiss albino mice and A549 lung cancer cells. B(a)P was orally supplemented (50 mg/kg body weight) twice a week for four weeks induction of lung cancer in mice. The lung weight, body weight, incidence of tumor, lipid peroxidation, carcinoembryonic antigen (CEA), enzymatic and nonenzymatic antioxidants (superoxide dismutase, GPx, glutathione, glutathione reductase, catalase, and glutathione S-transferase), serum marker enzymes (aryl hydroxylase, lactate dehydrogenase, 5'-nucleotidases, and γ-glutamyl transpeptidase), and inflammatory mediators (interleukin-1ß, interleukin-6, and tumor necrosis factor-α) were estimated. Moreover, a histopathological study of lung tissues was supported by the biochemical analysis. Furthermore, the anticancer activity of limonin on A549 cells was measured by cell viability, production of reactive oxygen species (ROS), apoptotic morphological changes by AO/EtBr staining. Additionally, the status of apoptosis protein (caspase-9 and -3) expressions was analyzed by the colorimetric analysis. B(a)P-induced mice showed increased lipid peroxidation, CEA, serum marker enzymes and inflammatory cytokines levels with simultaneously decreased in the nonenzymatic and enzymatic antioxidants levels. Limonin supplements significantly reverted back to all these changes in this manner, showing the efficiency of anticancer effect. Furthermore, our in vitro study also supported the anticancer effect of the treatment of limonin-enhanced apoptosis by loss of cell viability, improved ROS production, apoptotic morphological changes, and apoptosis protein expression were analyzed. Overall, these results suggest the anticancer potential of limonin against B(a)P-induced lung cancer in Swiss albino mice and A549 lung cancer cells.

Intestinal barrier dysfunction following traumatic brain injury.

Traumatic brain injury (TBI) can cause non-neurological injuries to other organs such as the intestine. Newer studies have shown that paracellular hyperpermeability is the basis of intestinal barrier dysfunction following TBI. Ischemia-reperfusion injury, inflammatory response, abnormal release of neurotransmitters and hormones, and malnutrition contribute to TBI-induced intestinal barrier dysfunction. Several interventions that may protect intestinal barrier function and promote the recovery of TBI have been proposed, but relevant studies are still limited. This review is to clarify the established mechanisms of intestinal barrier dysfunction following TBI and to describe the possible strategies to reduce or prevent intestinal barrier dysfunction.

[Rosiglitazone inhibited the transformation and synthesis of collagen in rats' embryonic lung fibroblasts through the modulation of plasminogen activator inhibitor-1].

OBJECTIVE: To investigate the role of plasminogen activator inhibitor-1 (PAI-1) in the inhibition of rosiglitazone on the transformation and collagen synthesis of rats' embryo lung fibroblasts and to examine the signal pathways in the process. METHODS: Fibroblasts from rats' embryo lung tissues were divided into 3 groups: Control, Rosiglitazone (Rosiglitazone 30 mmol/L) and PAI-1 (Rosiglitazone 30 mmol/L and PAI-1 20 mmol/L) groups. The fibroblasts were collected at 24, 48 and 72 h, and stored at -80 °C. RT-PCR was used to determine the expressions of collagen type-1 and type-3 at 24 h. Western blot analysis was used to determine the expressions of PAI-1, a-smooth muscle actin (α-SMA), p-AKT and p-ERK at 24, 48 and 72 h. RESULTS: There was a significant decrease in the protein expression of PAI-1 in the Rosiglitazone group (0.732±0.015, 0.583±0.005, 0.762±0.032) at 24, 48 and 72 h compared with the Control group (1.116±0.046). There was a significant increase in the protein expression in the PAI-1 group (0.923±0.042, 1.024±0.009, 1.070±0.011) compared with the Rosiglitazone group (F=78.609, P<0.01). The α-SMA protein expressions were significantly reduced in the Rosiglitazone group (0.209±0.012, 0.280±0.140, 0.254±0.025) compared with the Control group (0.340±0.026), while the expressions were significantly increased in the PAI-1 group (0.386±0.042, 0.400±0.037, 0.385±0.026) compared with the Rosiglitazone group (F=35.009, P<0.01). The collagen type-1 (1.065±0.004) and type-3 (1.282±0.001) mRNA expressions were significantly reduced in the Rosiglitazone group compared with the Control group (1.279±0.013, 1.690±0.005), while the expressions were significantly increased in the PAI-1 groups (1.390±0.029, 1.350±0.044) compared with the Rosiglitazone group (type-1: F=12.429, P<0.01; type-3: F=127.456, P<0.01). The expressions of p-AKT showed no differences among the 3 groups, but there were significant differences in the expressions of p-ERK in the Rosiglitazone group (0.288±0.010, 0.311±0.034, 0.336±0.038) compared with the Control group (0.506±0.032), and in PAI-1 groups (0.561±0.101, 0.448±0.022, 0.406±0.003) compared with the Rosiglitazone group (F=153.548, P<0.01). CONCLUSIONS: Rosiglitazone inhibits PAI-1 expression in fibroblasts from rats' embryo lung tissues and activates the fibrinolytic system. The up-regulation of PAI-1 expression alleviates the inhibition effect of rosiglitazone on the transformation and collagen synthesis of fibroblasts. The cross-talk between PAI-1 and ERK signal pathway may play an important role in the regulation of rosiglitazone on fibrosis.

The high expression of TC1 (C8orf4) was correlated with the expression of ß-catenin and cyclin D1 and the progression of squamous cell carcinomas of the tongue.

Thyroid cancer 1 (TC1, C8orf4) plays important roles in many signaling pathways, such as Wnt/ß-catenin signaling pathway, and is involved in the development of many cancers. The objective of this study was to examine the expression of TC1 and investigate the associations among TC1, ß-catenin, Chibby, cyclin D1, and the clinicopathological factors of oral tongue squamous cell carcinomas (OTSCCs). The expressions of TC1, ß-catenin, Chibby, and cyclin D1 were examined in 109 cases of OTSCCs using immunohistochemistry. The expression of TC1 was observed in all cases of OTSCCs but was negative or weak in normal squamous epithelial tissues of tongue. The high expression of TC1 was correlated with the advanced TNM stage (P = 0.042), the abnormal expression of ß-catenin (correlation coefficient = 0.314, P = 0.001) and the expression of cyclin D1 (correlation coefficient = 0.274, P = 0.006) in OTSCCs. But we did not find any associations between TC1 and Chibby. The abnormal expression of ß-catenin was correlated with the poor differentiation (P = 0.035), advanced TNM stage (P = 0.048) and the expression of cyclin D1 (correlation coefficient = 0.422, P < 0.001). In conclusion, the high expression of TC1 was common in OTSCCs and correlated with the expression of ß-catenin and cyclin D1 and the progression of OTSCCs. The high expression level of TC1 might promote the progression of OTSCCs by enhancing the activity of Wnt signaling pathway.

Extracellular vesicles in plant-microbe interactions: Recent advances and future directions.

Extracellular vesicles (EVs) are membrane-enclosed nanoparticles that have a crucial role in mediating intercellular communication in mammals by facilitating the transport of proteins and small RNAs. However, the study of plant EVs has been limited for a long time due to insufficient isolation and detection methods. Recent research has shown that both plants and plant pathogens can release EVs, which contain various bioactive molecules like proteins, metabolites, lipids, and small RNAs. These EVs play essential roles in plant-microbe interactions by transferring these bioactive molecules across different kingdoms. Additionally, it has been discovered that EVs may contribute to symbiotic communication between plants and pathogens. This review provides a comprehensive summary of the pivotal roles played by EVs in mediating interactions between plants and microbes, including pathogenic fungi, bacteria, viruses, and symbiotic pathogens. We highlight the potential of EVs in transferring immune signals between plant cells and facilitating the exchange of active substances between different species.

utPCR: A Strategy for the Highly Specific and Absolutely Quantitative Detection of Single Molecules within Only Minutes.

Bloodstream infection is a major health problem worldwide, with extremely high mortality. Detecting infection in the early stage is challenging due to the extremely low concentration of bacteria in the blood. Digital PCR provides unparalleled sensitivity and can achieve absolute quantification, but it is time-consuming. Moreover, the presence of unavoidable background signals in negative controls poses a significant challenge for single-molecule detection. Here, we propose a novel strategy called "Ultrafast flexible thin tube-based droplet digital PCR (utPCR)" that can shorten the digital PCR process from 2 h to only 5 min, with primer annealing/extension time reduced from minutes to only 5 s. Importantly, the ultrafast PCR eliminates nonspecific amplification and thus enables single-molecule detection. The utPCR enabled the sensitive detection and digital quantification of E. coli O157 in the high background of a 106-fold excess of E. coli K12 cells. Moreover, this method also displayed the potential to detect rare pathogens in blood samples, and the limit of detection (LOD) could be as low as 10 CFU per mL of blood without false positive results. Considered ultrafast (<5 min) and highly sensitive (single-molecule detection), the utPCR holds excellent prospects in the next generation of molecular diagnosis.

siRNA against plasminogen activator inhibitor-1 ameliorates bleomycin-induced lung fibrosis in rats.

AIM: Plasminogen activator inhibitor-1 (PAI-1) is involved in the progression of pulmonary fibrosis. The present study was undertaken to examine the effects on pulmonary fibrosis of silencing PAI-1 expression with small interfering RNA (siRNA) and to assess the possible underlying mechanisms. METHODS: Male Wistar rats were subjected to intratracheal injection of bleomycin (BLM, 5 mg/kg, 0.2 mL) to induce pulmonary fibrosis. Histopathological changes of lung tissue were examined with HE or Masson's trichrome staining. The expression levels of α-smooth muscle actin (α-SMA), collagen type-I and type-III, caspase-3, as well as p-ERK1/2 and PI3K/Akt in the lung tissue were evaluated using imunohistochemistry and Western blot analyses. The fibroblasts isolated from BLM-induced fibrotic lung tissue were cultured and transfected with pcDNA-PAI-1 or PAI-1siRNA. The expression level of PAI-1 in the fibroblasts was measured using real time RT-PCR and Western blot analysis. The fibroblast proliferation was evaluated using MTT assay. RESULTS: Intratracheal injection of PAI-1-siRNA (7.5 nmoL/0.2 mL) significantly alleviated alveolitis and collagen deposition, reduced the expression of PAI-1, α-SMA, collagen type-I and collagen type-III, and increased the expression of caspase-3 in BLM-induced fibrotic lung tissue. In consistence with the in vivo results, the proliferation of the cultured fibroblasts from BLM-induced fibrotic lung tissue was inhibited by transfection with PAI-1-siRNA, and accelerated by overexpression of PAI-1 by transfection with pcDNA-PAI-1. The expression of caspase-3 was increased as a result of PAI-1 siRNA transfection, and decreased after transfection with pcDNA-PAI-1. In addition, the levels of p-ERK1/2 and PI3K/Akt in the fibrogenic lung tissue were reduced after treatment with PAI-1siRNA. CONCLUSION: The data demonstrate that PAI-1 siRNA inhibits alveolitis and pulmonary fibrosis in BLM-treated rats via inhibiting the proliferation and promoting the apoptosis of fibroblasts. Suppression ERK and AKT signalling pathways might have at least partly contributed to this process. Targeting PAI-1 is a promising therapeutic strategy for pulmonary fibrosis.

Dual-color blending based visual LAMP for food allergen detection: A strategy with enlarged color variation range and contrast.

Food allergy has been a serious public health problem around the world. Its prevention relies heavily on the effective avoidance of any contaminated food, making clear and accurate detection very important. LAMP is one of the most potent methods for allergen rapid detection. However, its current colorimetric readouts usually have low color contrast and narrow color variation range. Thus, here we proposed a strategy based on color evolution to enlarge the variation range as well as the contrast to improve its suitability for naked-eye observation. By simply blending two commonly used color change processes during amplification, a wider color variation window, and a near contrast color change, purple-to-green with a hues difference of 10 were obtained. Three important allergens (walnuts, hazelnuts, and peanuts) were tested with a comparable sensitivity towards fluorescent real-time LAMP. Its feasibility for practical use has also been studied. This simple but effective strategy provides a new idea for the colorimetric detection of LAMP amplicons and can be applied to various fields.

Rapid, Visual, and Sequence-Specific Detection of Salmonella in Egg Liquid with vis-NEAA, a CRISPR/Cas12 Empowered New Strategy.

Salmonella is one of the main pathogenic factors that cause foodborne diseases. Rapid and accurate detection of Salmonella in food is of great importance to ensure food safety. Nicking enzyme-assisted amplification (NEAA) is one of the promising isothermal amplification methods finishing the in vitro amplification in ∼10 min; however, it suffers from nonspecific amplification a lot (∼70% products are noises). In this paper, we introduced CRISPR/Cas12a to specifically recognize the NEAA amplicons and transduce the signals into turned-on fluorescent visual readouts (vis-NEAA). Impressively, with this method, the high efficiency of NEAA has been taken great advantage and the nonspecific products were successfully bypassed at the same time. In comparison to NEAA-gel electrophoresis, vis-NEAA showed complete fidelity toward the presence of specific products, while for real-time PCR, it possesses equivalent sensitivity and specificity but saves ∼80% of the time. A level of 80 CFU/mL Salmonella in spiked eggs can be detected on-site in ∼20 min.

A predicting model of child-bearing-aged women' spontaneous abortion by co-infections of TORCH and reproductive tract.

To develop a predicting model of child-bearing-aged women' spontaneous abortion (SA) by co-infections of TORCH and reproductive tract, in order to provide a reference tool for accurately predicting the risk of SA and guide the early prevention, diagnosis and treatment of SA. A prospective cohort study was designed based on 218 958 child-bearing-aged women following up in Hebei province in China from 2010 to 2017. Multivariable logistic regression analysis was used to select candidate predictive variables. Fisher's discriminant analysis was performed to build a predictive model, and the validity of the model was evaluated. The incidence rate of SA was 2.4%. Multivariable logistic regression analysis showed that age (OR = 3.507), adverse pregnancy history (OR = 1.509), co-infections status of Candida and HBsAg (ORCandida positive×HBsAg negative  = 4.091, ORCandida negative×HBsAg positive  = 3.327, and ORCandida positive×HBsAg positive  = 13.762), and co-infections status of HBsAg, Rubella (IgG) and CMV (IgG) (ORHBs-Ag negative×Rubella (IgG) negative×CMV (IgG) positive  = 1.789, ORHBs-Ag positive×Rubella (IgG) positive×CMV (IgG) negative  = 3.809, and ORHBsAg positive×Rubella (IgG) positive×CMV (IgG) positive  = 11.919) were the independent predictors of SA. The total discriminant rate reached 91%, with 82% of the sensitivity and 91% of the specificity. The predicting model of child-bearing-aged women' SA by co-infections status has a good performance. The co-infection status of TORCH and reproductive tract are suggested to be considered in pre-pregnancy physical examination.

[The effect of plasminogen activator inhibitor-1 on embryonic lung fibroblasts from rats].

OBJECTIVE: To explore the effect of plasminogen activator inhibitor-1 (PAI-1) on the proliferation and conversion of rat embryonic lung fibroblasts and the synthesis of collagen, and therefore to explore the function of PAI-1 in pulmonary fibrosis. METHODS: The embryonic lung fibroblasts from pregnant Wistar rats were isolated and cultured in vitro. The reproduction rate of fibroblasts at 12 h, 24 h, and 48 h after being stimulated by PAI-1 with different concentrations (5, 10, 20, 40, 80, and 100 µg/L) was measured by MTT assay. After being stimulated by PAI-1 with the most suitable concentration (20 µg/L) for 48 h and 72 h, the expression of proliferating cell nuclear antigen (PCNA) was measured by immunocytochemical technique, and the mRNA expression of α-SMA and type-1 collagen at 24 h and 48 h was measured by real-time PCR. RESULTS: PAI-1 with different concentrations stimulated the proliferation of fibroblasts. The highest proliferation rate and absorbance in concentration of 20 µg/L and at 12 h were 62.6% and 0.573 ± 0.039. The comparison of different concentrations showed that the difference was significant (F = 111.112, P = 0.000). Therefore, 20 µg/L was selected as the most suitable concentration. Using immunocytochemical method, the optical density of PCNA at 48 h and 72 h were 3685 ± 686 and 2530 ± 477 after being stimulated with 20 µg/L PAI-1. The comparison showed significant difference (F = 7.85, P = 0.02). The expression of α-SMA increased (230 ± 11)% and (159 ± 9)% at 24 h and 48 h after being stimulated with 20 µg/L PAI-1, and the difference was significant (F = 39.92, P = 0.0003). The expression of type-1 collagen increased (92 ± 8)% and (65 ± 12)%, the difference being significant (F = 32.61, P = 0.0006). CONCLUSION: PAI-1 can promote the proliferation and conversion of fibroblasts and the synthesis of collagen, which may be involved in the pathogenesis of pulmonary interstitial fibrosis.

[Interventional effect of IL-17A on chronic obstructive pulmonary disease and its mechanism].

Objective: To investigate the intervention effects and mechanism of interleukin-17A (IL-17A) on chronic obstructive pulmonary disease (COPD). Methods: C57BL/6 mice were randomly divided into wild type blank control group, wild type COPD group and IL-7A knockout COPD group. Mice in wild type blank control group received no treatment, and mice in the other two groups were exposed to cigarette smoke to induce COPD (Cigarette: 1 cigarette / time, 4 times/day, 45 minutes/time; interval time: 1 hour; total intervention time: 90 days). Lung function of mice was assessed using animal pulmonary function machine. Bronchoalveolar lavage fluid (BALF) of mice was collected and BALF cell count and classification were determined. The lung tissue of mice was collected, the expression level of IL-17A in airway epithelium was determined by flow cytometry, and the levels of inflammatory factors in lung tissue were determined by enzyme-linked immunosorbent assay. The expression level of JNK/AP1 signaling pathway protein in mouse lung tissue was determined by Western blot. Results: Compared with the wild type blank control group mice, the wild type COPD group mice had significantly higher expression level of IL-17A, significantly lower peak inspiratory flow rate (PIF) and peak expiratory flow rate (PEF), significantly higher number of BALF neutrophils, eosinophils, lymphocytes and macrophage, significantly higher expression levels of CXC chemokine 1(CXCL1), CXC chemokine 2 (CXCL2), interleukin-1ß (IL-1ß) and interleukin-6 (IL-6), and significantly higher phosphorylation level of JNK, cJun and cFos and AP1 expression levels (P＜0.05). Compared with COPD mice, IL-17A expression level in airway epithelium of mice in IL-7A knockout COPD group was significantly lower, PIF and PEF were higher, the number of BALF neutrophils, eosinophils, lymphocytes and macrophage was significantly lower, the expression levels of CXCL1, CXCL2, IL-1ß and IL-6 in lung tissue were lower, and the phosphorylation levels of JNK, cJun and cFos and AP1 expression levels were significantly lower (P＜0.05). Conclusion: Cigarette smoke can induce the production of IL-17A and reduce (or inhibit) the production (or expression or secretion) of IL-17A in mouse airway epithelium, thus inhibiting the JNK/AP1 signaling pathway to reduce the airway inflammation and improve the lung function of COPD mice.

Thymic adenocarcinoma accompanied by type A thymoma and pulmonary minimally invasive adenocarcinoma and harboring distinct gene alterations: A case report.

RATIONALE: Thymic adenocarcinoma is an extremely rare thymic carcinoma. The exact genetic alteration associated with thymic adenocarcinoma is unclear. Here, we report a case of thymic adenocarcinoma accompanied by type A thymoma and pulmonary minimally invasive adenocarcinoma (MIA). PATIENT CONCERNS: A 53-year-old woman presented with multiple nodules in the mediastinum and lung. Thoracic computed tomography revealed nodules in the anterior superior mediastinum and anterior mediastinum near the right pericardium and ground-glass opacity (GGO) in the right superior lobe of the lung. DIAGNOSIS: The tumor in the anterior superior mediastinum was diagnosed as primary thymic papillary adenocarcinoma. The tumor in the anterior mediastinum near the right pericardium was diagnosed as type A thymoma. The GGO of the right superior lobe of the lung was diagnosed as a MIA. INTERVENTION: The patient underwent thoracoscopic mediastinal tumor resection and partial lobectomy in our hospital. OUTCOMES: The postoperative course was uneventful. The patient is alive and free of the disease for 22 months after diagnosis. LESSONS: Thyroid transcription factor 1 (TTF-1) was positive in this case of thymic adenocarcinoma, which indicated that a thymic adenocarcinoma with TTF-1-positive may not necessarily be a metastasis of lung or thyroid adenocarcinoma. The positive staining of CD5 and CD117 can help us to confirm the thymic origin. Molecular genetic analysis indicated that these tumors harbored different mutations. The thymic adenocarcinoma and type A thymoma both had the mutation of KMT2A, but the mutation sites were different. KMT2A mutation may be a common genetic change in thymic tumorigenesis. The genetic alterations disclosed in this study will help expand the understanding of thymic tumors.

DEK is highly expressed in breast cancer and is associated with malignant phenotype and progression.

DEK proto-oncogene (DEK) has been demonstrated as an oncogene and is associated with the development of many types of tumor; however, the expression and role of DEK in breast cancer remain unknown. The present study aimed to determine the role of DEK in the progression of breast cancer. The expression of DEK in 110 breast cancer tissues and 50 adjacent normal breast tissues was examined using immunohistochemistry. Furthermore, DEK expression was upregulated by DEK transfection or downregulated by DEK shRNA interference in MCF7 cells. Proliferative and invasive abilities were examined in MCF7 cells using MTT assay, colony-formation assay and transwell invasion assays. The results demonstrated that DEK expression level was significantly increased in breast cancer tissues compared with normal breast tissues. Furthermore, high DEK expression was associated with high histological grade, lymph node metastasis, advanced Tumor-Node-Metastasis stage and high Ki-67 index; however, DEK expression was not associated with the expression level of estrogen receptor, progesterone receptor, and human epidermal growth factor receptor 2. High DEK expression indicated poor prognosis in patients with breast cancer. DEK overexpression upregulated the protein expression of ß-catenin and Wnt and increased the proliferative and invasive abilities of breast cancer cells. DEK downregulation had the opposite effect. Taken together, the results from the present study demonstrated that high expression of DEK was common in patients with breast cancer and was associated with progression of the disease and poor prognosis, and that DEK overexpression promoted the proliferative and invasive abilities of breast cancer cells.

Protective effect of rutin against bleomycin induced lung fibrosis: Involvement of TGF-ß1/α-SMA/Col I and III pathway.

Lung fibrosis is a progressive fatal lung disorder with significantly high mortality rates. Bleomycin (BLM) is one of the most commonly used chemotherapeutic agents for the treatment of several carcinomas. The most severe adverse effect of BLM is lung toxicity; therefore, BLM has been repeatedly reported to be considered amongst the most widely used agents for the induction of experimental lung fibrosis. In the current study, rutin has been investigated for its ability to ameliorate BLM-induced pulmonary fibrosis. BLM was instilled intratracheally and rutin was administered orally (50 and 100 mg/kg) for 3 weeks. Rutin significantly decreased lung/body weight index, bronchoalveolar lavage fluid lactate dehydrogenase activity, total cell count, macrophages, and lymphocyte counts. Rutin significantly decreased lung malondialdehyde content, increased lung glutathione content, superoxide dismutase activity, serum total antioxidant capacity, and decreased lung nitric oxide content. Moreover, rutin reduced expressions of transforming growth factor beta 1 and other fibrosis-related biomarkers (Col I, Col III, and α-SMA). In addition, rutin significantly ameliorated histological changes and prevented collagen deposition with the paralleled decrease in lung hydroxyproline content. In conclusion, rutin can be proposed to be a potential therapeutic agent for the management of lung fibrosis.

Primary thymic carcinoma with adenoid cystic carcinoma-like features: A case report and literature review.

RATIONALE: Thymic carcinoma with adenoid cystic carcinoma-like features is a special subtype of thymic adenocarcinoma, and the occurrence of this condition is extremely rare. Herein, we report a case of primary thymic carcinoma with adenoid cystic carcinoma-like features in a young man. PATIENT CONCERNS: A 38-year-old man had an incidental finding of space-occupying lesion in the anterior mediastinum during a routine health examination. The patient complained of occasional mild chest tightness during hot weather but had no obvious cough, sputum, chest pain, or fever. Contrast-enhanced computed tomography scan of the chest revealed a space-occupying lesion in the anterior mediastinum, which is likely benign. DIAGNOSIS: The lesion was diagnosed as a primary thymic carcinoma with adenoid cystic carcinoma-like features. INTERVENTION: The patient underwent thoracoscopic resection of left anterior mediastinal mass and enlarged resection of thymectomy and mediastinal fat in our hospital. OUTCOMES: The postoperative course was uneventful. LESSONS: The tissue characteristic of this tumor was extremely similar to that of adenoid cystic carcinoma. A precise pathological examination is extremely important to prevent misdiagnoses of the lesion as adenoid cystic carcinoma or other thymic tumors. Immunohistochemical staining is extremely useful for the pathological and differential diagnoses of this tumor.

DEK promotes the proliferation and invasion of lung cancers and indicates poor prognosis in lung adenocarcinomas.

DEK has been revealed to be overexpressed in many cancers and associated with cancer progression. The aim of the present study was to elucidate the role of DEK with a specific focus on its underlying mechanism in lung cancers. DEK expression in lung cancers and normal lung tissues and the correlations between DEK expression and clinicopathological parameters of lung cancers were investigated using the data from The Cancer Genome Atlas (TCGA). DEK expression was upregulated by DEK transfection or downregulated by DEK shRNA interference in A549 and H1299 cells. The effects of DEK on the Wnt signaling pathway and epithelial­mesenchymal transition (EMT) were examined using western blotting. Proliferative and invasive abilities were observed in A549 and H1299 cells treated with DEK using an MTT assay, colony formation assay, and Transwell migration and invasion assays. The expression of DEK was higher in lung cancer tissues than that in normal lung tissues. DEK expression was positively correlated with the expression of epidermal growth factor receptor (EGFR) and KRAS in lung adenocarcinomas. High expression of DEK indicated poor prognosis in lung adenocarcinomas (P=0.018). Enhanced expression of DEK upregulated the levels of active­ß­catenin and Wnt target genes, such as cyclin D1, c­Myc and MMP7 and increased the proliferative and invasive abilities of lung cancer cells. Enhanced expression of DEK in A549 and H1299 cells also increased the levels of EGFR, KRAS, vimentin, Snail, and N­cadherin, and decreased the level of E­cadherin. The opposite results were obtained with knockdown of DEK expression. DEK was highly expressed in lung cancers and indicated poor prognosis in lung adenocarcinomas. DEK expression activated the Wnt signaling pathway and EMT process and promoted the proliferation and invasion of lung cancers.

Genetic association of ADAM33 polymorphisms with childhood asthma in Chinese Han population: A case-control study.

To explore the association of a disintegrin and metalloprotease 33 (ADAM33) polymorphisms with childhood asthma susceptibility, we conducted this case-control study.In this case-control study, we selected 96 asthma children and 86 healthy children to conduct the genotyping of ADAM33 polymorphisms through polymerase chain reaction-direct sequencing (PCR-DS). Hardy-Weinberg equilibrium (HWE) status in the control group was detected adopting chi-square test. Frequency differences of genotypes, alleles, and haplotypes were compared by chi-square test between the case and control groups. Linkage disequilibrium (LD) between polymorphisms was checked using Haploview software. Association intensity of the polymorphisms with the disease risk was assessed by odds ratio (OR) and 95% confidence interval (95%CI).The frequency of rs678881 GA genotype was obviously higher in cases than in controls (P = .03) and the carriage of this genotype conferred higher risk of asthma among children than GG genotype (OR = 2.03, 95%CI = 1.05-3.91). However, neither rs2280089 nor rs2853209 polymorphism was significantly associated with the risk of childhood asthma. Strong LD was found among rs678881, rs2280089 and rs2853209, and haplotype GGT was distinctly associated with the risk of asthma in children (OR = 0.28, 95%CI = 0.13-0.57).ADAM33 rs678881 polymorphism is significantly correlated with increased susceptibility to asthma in Chinese Han children. Besides, haplotype GGT among the 3 polymorphisms was obviously associated with decreased risk of childhood asthma.